SSAT MRNA TRANSLATION REPRESSION AND ACTIVATION

The invention provides an isolated nucleic acid having a sequence encoding a spermidine/spermine acetyltransferase ("SSAT"), wherein translation of an mRNA comprising the encoded SSAT has increased basal translation and increased stimulated translation, compared to a wild-type mRNA encoding SSAT. Methods of use for the nucleic acid are also provided. Methods and compositions are also provided for reducing ischemia-reperfusion injury in organs or tissue for transplantation.

METHODS OF ALTERING SEED WEIGHT AND SEED OIL CONTENT BY MANIPULATING ALPHA-CARBOXYLTRANSFERASE (A-CT) ACTIVITY VIA CARBOXYLTRANSFERASE INTERACTOR (CTI) PROTEIN EXPRESSION

A method of altering fatty acid and/or triacylglycerol production in plants and/or algae is provided. The method includes altering activity levels of alpha-carboxyltransferase (a-CT), a catalytic subunit of acetyl-CoA carboxylase (ACCase) by modulating an expression of at least one carboxyltransferase interaction (CTI) gene encoding at least one CTI protein.

SSAT mRNA TRANSLATION REPRESSION AND ACTIVATION

The invention provides an isolated nucleic acid having a sequence encoding a spermidine/spermine acetyltransferase (SSAT), wherein translation of an mRNA comprising the encoded SSAT has increased basal translation and increased stimulated translation, compared to a wild-type mRNA encoding SSAT. Methods of use for the nucleic acid are also provided. Methods and compositions are also provided for reducing ischemia-reperfusion injury in organs or tissue for transplantation.

SSAT MRNA TRANSLATION REPRESSION AND ACTIVATION

The invention provides an isolated nucleic acid having a sequence encoding a spermidine/spermine acetyltransferase ("SSAT"), wherein translation of an mRNA comprising the encoded SSAT has increased basal translation and increased stimulated translation, compared to a wild-type mRNA encoding SSAT. Methods of use for the nucleic acid are also provided. Methods and compositions are also provided for reducing ischemia-reperfusion injury in organs or tissue for transplantation.

SSAT mRNA translation repression and activation

The invention provides an isolated nucleic acid having a sequence encoding a spermidine/spermine acetyltransferase (“SSAT”), wherein translation of an mRNA comprising the encoded SSAT has increased basal translation and increased stimulated translation, compared to a wild-type mRNA encoding SSAT. Methods of use for the nucleic acid are also provided. Methods and compositions are also provided for reducing ischemia-reperfusion injury in organs or tissue for transplantation.

MICROORGANISMS HAVING PUTRESCINE PRODUCTIVITY AND PROCESS FOR PRODUCING PUTRESCINE USING THE SAME

The present invention relates to a recombinant microorganism capable of producing putrescine, in which the microorganism is modified to have enhanced NCgl2522 activity, thereby producing putrescine in a high yield, and a method for producing putrescine using the microorganism. 1. A microorganism having putrescine productivity , which is modified to have enhanced activity of a protein having an amino acid sequence represented by SEQ ID NO: 21 or 23.2. The microorganism having putrescine productivity according to claim 1 , wherein the microorganism is further modified to have weakened activities of ornithine carbamoyltransferase (ArgF) and a protein (NCgl1221) involved in glutamate export claim 1 , compared to the endogenous activities claim 1 , and to have enhanced ornithine decarboxylase (ODC) activity.3. The microorganism having putrescine productivity according to claim 2 , wherein the ornithine carbamoyltransferase (ArgF) has an amino acid sequence represented by SEQ ID NO: 29 claim 2 , the protein (NCgl1221) involved in glutamate export has an amino acid sequence represented by SEQ ID NO: 30 claim 2 , and the ornithine decarboxylase (ODC) has an amino acid sequence represented by SEQ ID NO: 33.4. The microorganism having putrescine productivity according to claim 1 , wherein the microorganism is further modified to have enhanced activities of acetyl-gamma-glutamyl-phosphate reductase (ArgC) claim 1 , acetylglutamate synthase or ornithine acetyltransferase (ArgJ) claim 1 , acetylglutamate kinase (ArgB) claim 1 , and acetylornithine aminotransferase (ArgD) claim 1 , compared to the endogenous activities.5. The microorganism having putrescine productivity according to claim 4 , wherein the acetyl-gamma-glutamyl-phosphate reductase (ArgC) claim 4 , acetylglutamate synthase or ornithine acetyltransferase (ArgJ) claim 4 , acetyl glutamate kinase (ArgB) claim 4 , and acetylornithine aminotransferase (ArgD) have amino acid sequences represented by SEQ ID NOs: 25 claim 4 ...

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification conferring stability to the RNA, as well as related methods, are disclosed. 120-. (canceled)21. A method of in vivo protein expression , comprising administration of a composition comprising at least one mRNA molecule encoding a protein of interest and a 5′ untranslated region (UTR) comprising SEQ ID NO: 1 , wherein SEQ ID NO: 1 enhances the expression of the protein of interest.22. The method of claim 21 , wherein said mRNA molecule further comprises a poly A tail.23. The method of claim 21 , wherein said mRNA molecule further comprises a Cap1 structure.24. The method of claim 21 , wherein said mRNA molecule further comprises a 3′ untranslated region (UTR).25. The method of claim 24 , wherein said 3′ UTR comprises a sequence encoding human growth hormone (hGH).26. The method of claim 25 , wherein said sequence encoding human growth hormone (hGH) comprises SEQ ID NO: 3.27. The method of claim 21 , wherein the protein of interest is ornithine carbamoyltransferase.28. The method of claim 21 , wherein the protein of interest is alpha galactosidase.29. The method of claim 21 , wherein the protein of interest is erythropoietin.30. The method of claim 21 , wherein the protein is expressed in a target cell selected from hepatocytes claim 21 , epithelial cells claim 21 , hematopoietic cells claim 21 , endothelial cells claim 21 , lung cells claim 21 , bone cells claim 21 , stem cells claim 21 , mesenchymal cells claim 21 , neural cells claim 21 , cardiac cells claim 21 , adipocytes claim 21 , vascular smooth muscle cells claim 21 , cardiomyocytes claim 21 , skeletal muscle cells claim 21 , beta cells claim 21 , pituitary cells claim 21 , synovial lining cells claim 21 , ovarian cells claim 21 , testicular cells claim 21 , fibroblasts claim 21 , B cells claim 21 , T cells claim 21 , reticulocytes claim 21 , leukocytes claim ...

Cleavable Lipids

Disclosed herein are novel compounds, pharmaceutical compositions comprising such compounds and related methods of their use. The compounds described herein are useful, e.g., as liposomal delivery vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and subsequent iransfection of said target cells, and in certain embodiments are characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids. 2. (canceled)3. The nanoparticle of claim 1 , wherein Ris imidazole.4. The nanoparticle of claim 1 , wherein{'sub': '1', 'Ris imidazole;'}andn is 1.5. The nanoparticle of claim 1 , wherein Ris guanidinium.6. The nanoparticle-of claim 1 , wherein{'sub': '1', 'Ris guanidinium;'}andn is 1.723.-. (canceled)2629.-. (canceled)30. The nanoparticle of claim 1 , further comprising one or more compounds selected from the group consisting of a cationic lipid claim 1 , a PEG-modified lipid claim 1 , a non-cationic lipid and a helper lipid.31. (canceled)32. The nanoparticle of claim 1 , wherein one or more of the polynucleotides comprises a chemical modification.33. The nanoparticle of claim 1 , wherein the one or more polynucleotides is selected from the group consisting of an antisense oligonucleotide claim 1 , siRNA claim 1 , miRNA claim 1 , snRNA claim 1 , snoRNA and combinations thereof.34. (canceled)35. The nanoparticle of claim 1 , wherein the one or more polynucleotides comprise DNA.36. The nanoparticle of claim 1 , wherein the one or more polynucleotides comprise RNA.37. (canceled)38. The nanoparticle of claim 36 , wherein the RNA encodes an enzyme.39. The nanoparticle of claim 38 , wherein the enzyme is selected from the group consisting of agalsidase alfa claim 38 , alpha-L-iduronidase claim 38 , iduronate-2-sulfatase claim 38 , N-acetylglucosamine-1-phosphate transferase claim 38 , N-acetylglucosaminidase claim 38 , alpha-glucosaminide acetyltransferase claim 38 , N- ...

Messenger RNA Therapy for the Treatment of Ornithine Transcarbamylase Deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase (OTC) deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase (OTC) protein at an effective dose and an administration interval such that at least one symptom or feature of the OTC deficiency is reduced in intensity, severity, or frequency or has delayed onset. In some embodiments, the mRNA is encapsulated in a liposome comprising one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids. 1. A method of treating ornithine transcarbamylase (OTC) deficiency , comprising administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbyamylase (OTC) protein at an effective dose and an administration interval such that at least one symptom or feature of the OTC deficiency is reduced in intensity , severity , or frequency or has delayed onset.2. The method of claim 1 , wherein the mRNA encoding the OTC protein is codon optimized.3. The method of claim 2 , wherein the mRNA encoding the OTC protein comprises a polynucleotide sequence at least 70% claim 2 , 75% claim 2 , 80% claim 2 , 85% claim 2 , 90% claim 2 , or 95% identical to SEQ ID NO: 3 claim 2 , SEQ ID NO: 6 claim 2 , SEQ ID NO: 7 claim 2 , SEQ ID NO: 8 claim 2 , SEQ ID NO: 9 or SEQ ID NO: 10.4. The method of claim 3 , wherein the mRNA encoding the OTC protein comprises SEQ ID NO: 3 claim 3 , SEQ ID NO:6 claim 3 , SEQ ID NO:7 claim 3 , SEQ ID NO:8 claim 3 , SEQ ID NO:9 or SEQ ID NO:10.5. The method of claim 4 , wherein the mRNA encoding the OTC protein comprises SEQ ID NO: 7.6. The method of claim 2 , wherein the mRNA encoding the OTC protein is not SEQ ID NO: 1.73. The method of claim any one of the preceding claims claim 2 , wherein the mRNA further comprises the 5′ untranslated region (UTR) sequence of SEQ ID ...

Stable gene transfer to proliferating cells

Provided herein are methods for facilitating or inducing stable transgene integration and expression in a proliferating cell, comprising administering to the cell (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences, which sequences are in turn flanked by AAV-derived inverted terminal repeat regions, and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell. Also provide are methods and transgene delivery systems for the treatment or prevention of diseases affecting, associated with or characterised by proliferating cells.

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Non-viral delivery systems comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A composition comprising a non-viral carrier and a nucleic acid molecule comprising an engineered sequence encoding human ornithine transcarbamylase (hOTCase) and expression control sequences which direct expression of hOTCase in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over at least the mature hOTC sequence of SEQ ID NO: 1 , wherein said hOTC nucleic acid sequence is selected from the nucleic acid sequence comprising SEQ ID NO: 5 or a nucleic acid sequence at least about 96 to about 99% identical thereto , a nucleic acid sequence selected from SEQ ID NO: 9 , or a nucleic acid sequence at least about 96 to about 99% identical thereto , a nucleic acid sequence comprising SEQ ID NO: 11 or a nucleic acid sequence at least about 96 to about 99% identical thereto , or a nucleic acid sequence comprising SEQ ID NO: 13 or a nucleic acid sequence at least about 96 to about 99% identical thereto.2. The composition according to claim 1 , wherein the hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 4.3. The composition according to claim 1 , wherein the hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 3.4. The composition according to claim 1 , wherein the hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 8.5. The composition according to claim 1 , wherein the hOTC is an RNA sequence of SEQ ID NO: 10.6. The composition according to claim 1 , wherein the hOTC is an RNA sequence of SEQ ID NO: 12.7. The composition according to claim 1 , wherein the hOTC is a chimeric OTC comprises a ...

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification 5 conferring stability to the RNA, as well as related methods, are disclosed. 1. A composition for modulating the expression of a protein in a target cell , wherein said composition comprises at least one RNA molecule and a transfer vehicle and wherein the RNA comprises at least one modification which confers stability to the RNA.2. The composition of claim 1 , wherein the RNA molecule is selected from the group consisting of mRNA claim 1 , miRNA claim 1 , snRNA claim 1 , and snoRNA.3. The composition of claim 1 , wherein the RNA molecule comprises more than one modification which confers stability to the RNA molecule.4. The composition of claim 1 , wherein the RNA molecule comprises a modification of the 5′ untranslated region of said RNA molecule.5. The composition of claim 4 , wherein said modification comprises a partial sequence of a CMV immediate-early 1 (IE 1) gene.6. The composition of claim 5 , wherein said partial sequence of the CMV immediate-early 1 (IEI) gene comprises SEQ ID NO: 2 or SEQ ID NO: 1.7. The composition of claim 4 , wherein said modification comprises the inclusion of a poly A tail.8. The composition of claim 4 , wherein said modification comprises the inclusion of a Cap 1 structure.9. The composition of claim 1 , wherein the RNA molecule comprises a modification of the 3′ untranslated of said RNA molecule.10. The composition of claim 9 , wherein said modification comprises the inclusion of a sequence encoding human growth hormone (hGH).11. The composition of claim 10 , wherein said sequence encoding human growth hormone (hGH) comprises SEQ ID NO: 3.12. The composition of claim 9 , wherein said modification comprises the inclusion of a poly A tail.13. The composition of claim 1 , wherein the RNA encodes ornithine carbamoyl transferase.14. The composition of claim 1 , wherein the RNA encodes ...

COMPOSITIONS AND METHODS FOR FACILITATING DELIVERY OF SYNTHETIC NUCLEIC ACIDS TO CELLS

Provided herein are compositions and methods for facilitating or enhancing delivery of nucleic acids, such as synthetic mRNAs, into cells or tissues. Such compositions and methods may include use of a targeting moiety-conjugated, such as an N-acetylgalactosamine (GalNAc)-conjugated, oligonucleotide to facilitate or enhance delivery. 1. A composition comprising:(a) an oligonucleotide of up to 50 nucleotides in length covalently linked to a targeting moiety; and(b) a synthetic RNA comprising at least one binding region that is complementary to a contiguous stretch of at least 5 nucleotides of the oligonucleotide.2. The composition of claim 1 , wherein the synthetic RNA comprises at least two copies of the binding region.3. The composition of claim 1 , wherein the synthetic RNA comprises at least three copies of the binding region.4. The composition of claim 1 , wherein the synthetic RNA comprises at least four copies of the binding region.5. The composition of claim 1 , wherein the synthetic RNA comprises at least five copies of the binding region.6. The composition of claim 1 , wherein the synthetic RNA comprises at least six copies of the binding region.7. The composition of claim 1 , wherein the synthetic RNA comprises at least seven copies of the binding region.8. The composition of any one of to claim 1 , wherein the copies of the binding region are separated from one another by a spacer region comprising at least one nucleotide.9. The composition of claim 8 , wherein each spacer region between each copy of the binding region is independently between 3 and 24 nucleotides in length.10. The composition of claim 8 , wherein each spacer region between each copy of the binding region is independently 3 claim 8 , 6 claim 8 , 12 or 24 nucleotides in length.11. The composition of any one of to claim 8 , wherein the synthetic RNA is a synthetic mRNA.12. The composition of claim 11 , wherein the at least one binding region is located in an untranslated region (UTR) of the ...

Microorganism for producing putrescine or ornithine and process for producing putrescine or ornithine using them

Disclosed is a modified microorganism producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same.

MESSENGER UNA MOLECULES AND USES THEREOF

This invention provides a range of translatable messenger UNA (mUNA) molecules. The mUNA molecules can be translated in vitro and in vivo to provide an active polypeptide or protein, or to provide an immunization agent or vaccine component. The mUNA molecules can be used as an active agent to express an active polypeptide or protein in cells or subjects. Among other things, the mUNA molecules are useful in methods for treating rare diseases. 1. A mUNA molecule , comprising one or more UNA monomers , and comprising nucleic acid monomers , wherein the mUNA molecule is translatable to express a polypeptide or protein.2. The molecule of claim 1 , wherein the molecule comprises from 200 to 12 claim 1 ,000 monomers.3. The molecule of claim 1 , wherein the molecule comprises from 200 to 4 claim 1 ,000 monomers.4. The molecule of claim 1 , wherein the molecule comprises from 1 to 8 claim 1 ,000 UNA monomers.5. The molecule of claim 1 , wherein the molecule comprises from 1 to 100 UNA monomers.6. The molecule of claim 1 , wherein the molecule comprises from 1 to 20 UNA monomers.7. The molecule of claim 1 , wherein the molecule comprises one or more modified nucleic acid nucleotides claim 1 , or one or more chemically-modified nucleic acid nucleotides.8. The molecule of claim 1 , wherein the molecule comprises a 5′ cap claim 1 , a 5′ untranslated region of monomers claim 1 , a coding region of monomers claim 1 , a 3′ untranslated region of monomers claim 1 , and a tail region of monomers.9. The molecule of claim 8 , wherein the molecule comprises a translation enhancer in a 5′ or 3′ untranslated region.10. The molecule of claim 1 , wherein the molecule is translatable in vivo.11. The molecule of claim 1 , wherein the molecule is translatable in vitro.12. The molecule of claim 1 , wherein the molecule is translatable in a mammalian cell.13. The molecule of claim 1 , wherein the molecule is translatable in a human in vivo.14. The molecule of claim 1 , wherein a translation ...

Compositions useful in treatment of ornithine transcarbamylase (otc) deficiency

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis cirrhosis in OTCD patients by administering hOTC.

Composition and Methods for Treatment of Ornithine Transcarbamylase Deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase protein at a low dose and at an administration interval such that at least one symptom or feature of the OTC deficiency is reduced. 1. A method of treating ornithine transcarbamylase (OTC) deficiency in a human , comprising administering to a human in need of treatment a pharmaceutical composition comprising an mRNA encoding an ornithine transcarbamylase protein formulated in a lipid nanoparticle ,wherein the mRNA encoding the ornithine transcarbamylase protein is administered at a therapeutic low dose of 0.5 mg/kg or less of mRNA at a dosing interval of once every two weeks or a longer dosing internal for a period sufficient to treat at least one symptom or reduce the level of a biomarker associated with ornithine transcarbamylase deficiency in the human relative to a control.2. The method of claim 1 , wherein the therapeutic low dose is 0.4 mg/kg or less claim 1 , or 0.3 mg/kg or less claim 1 , or 0.2 mg/kg or less claim 1 , or 0.15 mg/kg less claim 1 , or 0.10 mg/kg or less claim 1 , or 0.05 mg/kg or less claim 1 , or 0.01 mg/kg or less claim 1 , of mRNA encoding ornithine transcarbamylase protein.37-. (canceled)8. The method of claim 1 , wherein the longer dosing interval is once every three weeks or longer claim 1 , or once every 4 weeks or longer.9. The method of claim 1 , wherein the mRNA is codon optimized.10. The method of claim 1 , wherein the symptom comprises hyperammonemia.11. The method of claim 1 , wherein the biomarker is selected from the group consisting of: high plasma ammonia level claim 1 , high tissue ammonia level claim 1 , urinary orotic acid claim 1 , citrulline claim 1 , serum glutamate claim 1 , brain myoinositol claim 1 , serum amino acids claim 1 , and combination thereof.1213-. (canceled)14. A method ...

MULTIMERIC CODING NUCLEIC ACID AND USES THEREOF

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends. 1. A multimeric coding nucleic acid (MCNA) comprising two polynucleotides linked via 3′ ends via an oligonucleotide bridge comprising a stable 3-3′ inverted phosphodiester linkage such that the multimeric coding nucleic acid compound comprises two or more 5′ ends , andwherein at least one of the two polynucleotides is an encoding polynucleotide.24.-. (canceled)5. The MCNA of claim 1 , wherein the at least one encoding polynucleotide encodes a protein of interest.6. The MCNA of claim 5 , wherein the two polynucleotides are encoding polynucleotides claim 5 , and each of the two encoding polynucleotides encodes the same protein.7. The MCNA of claim 5 , wherein the two polynucleotides are encoding polynucleotides claim 5 , and each of the two encoding polynucleotides encodes a distinct protein.811.-. (canceled)12. The MCNA of claim 1 , wherein the at least one encoding polynucleotide comprises a 3′ UTR.13. (canceled)14. The MCNA of claim 12 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.1526.-. (canceled)27. The MCNA of claim 1 , wherein the nucleosides comprising the oligonucleotide bridge are selected from the group consisting of 2′-OMe-A claim 1 , 2′-OMe-G claim 1 , 2′-OMe-C claim 1 , 2′-OMe-U claim 1 , 2′-F-A claim 1 , 2′-F-G claim 1 , 2′-F-C claim 1 , 2′-F-U claim 1 , LNA-A claim 1 , LNA-G claim 1 , LNA-C claim 1 , LNA-U claim 1 , N6-methyl-adenosine claim 1 , 2-thiouridine (2sU) claim 1 , 5-methyl-cytidine (5mC) claim 1 , pseudouridine (ψU) claim 1 , and 1-methyl-pseudouridine.2830.-. (canceled)31. The MCNA of claim 1 , wherein the at least one encoding polynucleotide ...

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A recombinant viral vector comprising a nucleic acid sequence encoding human ornithine transcarbamylase (hOTC) and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over the mature sequence or full length hOTC of SEQ ID NO: 1 , and expresses a functional hOTC , wherein said hOTC nucleic acid sequence is SEQ ID NO: 9 or a nucleic acid sequence at least 96 to 99% identical thereto.2. A recombinant viral vector comprising a nucleic acid sequence encoding hOTC and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence expresses a functional hOTC , wherein said hOTC nucleic acid sequence is SEQ ID NO: 3. This is a continuation of U.S. patent application Ser. No. 15/122,853, filed Aug. 31, 2016, which is a national stage application under 35 U.S.C. 371 of PCT/US2015/019513, filed on Mar. 9, 2015, now expired, which claims the benefit of U.S. Patent Application No. 61/950,157, filed Mar. 9, 2014, now expired. These applications are incorporated by reference in their entirety.This invention was made with support under grant Nos. P01-HD057247, P01-HL059407, and P30-DK047757 awarded by the National Institutes of Health. The US government has certain rights in the invention.Ornithine transcarbamylase (OTC) deficiency accounts for nearly half of all cases of inborn errors of urea synthesis, with a prevalence estimated to be at least 1 in 15,000. Urea cycle defects put patients at risk of life threatening ...

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification 5 conferring stability to the RNA, as well as related methods, are disclosed. 120-. (canceled)21. A method of treating a urea cycle disorder comprisingadministering to a subject suffering from or susceptible to a urea cycle disorder, a composition comprising an mRNA encoding an enzyme, encapsulated within a liposome;wherein the enzyme is selected from ornithine transcarbamylase (OTC), carbamyl phosphate synthetase (CPS), argininosuccinate synthetase 1 (ASS1) argininosuccinate lyase (ASL), or arginase (ARG).22. The method of claim 21 , wherein the enzyme is ASS1.23. The method of claim 21 , wherein the mRNA is administered systemically.24. The method of claim 21 , wherein the composition is administered intravenously.25. The method of claim 21 , wherein the composition is administered bi-weekly.26. The method of claim 21 , wherein the composition is administered monthly.27. The method of claim 21 , wherein the administering of the composition results in increased expression of the enzyme encoded by the mRNA in the liver.28. The method of claim 21 , wherein the mRNA comprises a modification to enhance stability.29. The method of claim 28 , wherein the modification is selected from a group consisting of a modified nucleotide claim 28 , an alteration to the 5′ or 3′ untranslated region (UTR) claim 28 , a cap structure and a poly A tail.30. The method of claim 29 , wherein the modification comprises a pseudouridine.31. The method of claim 29 , wherein the modification comprises a partial sequence of a CMV immediate-early 1 (IE1) gene.32. The method of claim 31 , wherein said partial sequence of the CMV immediate-early (IE1) gene comprises SEQ ID NO: 1 claim 31 , wherein SEQ ID NO: 1 enhances the expression of the protein of interest. The subject application claims the benefit of U.S. Provisional Application Ser. No. 61/ ...

Disease model pig exhibiting stable phenotype, and production method thereof

Disease model pigs produced by nuclear transplantation, disease model pigs exhibiting stable phenotypes and production methods thereof are provided. Chimeric pigs for producing disease model pigs exhibiting stable phenotypes, genital glands thereof, and germ cells thereof are also provided. A method for producing a genetically modified disease model pig, includes: (a) transplanting a nucleus of a genetically modified cell into cytoplasm of an egg; (b) developing an obtained clonal embryo in a womb of a female pig to obtain an offspring; and mating the obtained offspring or having the offspring undergo sexual reproduction to further obtain the genetically modified offspring as a disease model pig.

L-ORNITHINE PRODUCTION IN EUKARYOTIC CELLS

The present invention relates to the provision of genetically modified microbial cells, such as yeast cells with an improved ability for producing L-ornithine and its derivatives. Overproduction of L-ornithine is obtained in the first place by the down-regulation or attenuation of specially selected genes, wherein said genes encode enzymes involved in the L-ornithine consumption and/or degradation pathways. Further L-ornithine production ability is improved by down-regulation, attenuation, deletion or overexpression of specially selected genes, wherein said genes encode enzymes and/or proteins involved in the L-ornithine ‘acetylated derivatives cycle’, L-glutamate synthesis pathways, subcellular trafficking, TCA cycle, pyruvate carboxylation pathway, respiratory electron-transport chain, and the carbon substrates' assimilation machinery. The invention additionally provides a method to produce L-ornithine with said modified eukaryotic cells. 139-. (canceled)40. A genetically modified eukaryotic cell modified for enhanced L-ornithine biosynthesis from α-ketoglutarate , wherein (a) a gene encoding a cytosolic glutamate N-acetyltransferase,', '(b) a gene encoding a cytosolic N-acetylglutamate kinase,', '(c) a gene encoding a cytosolic N-acetyl-gamma-glutamyl-phosphate reductase,', '(d) a gene encoding a cytosolic acetylornithine aminotransferase and', '(e) gene encoding a cytosolic ornithine acetyltransferase; or, '(1) the L-ornithine biosynthesis from α-ketoglutarate is cytosolic and the eukaryotic cell is modified to comprise at least one of the genes selected from a group consisting of'}(2) the eukaryotic cell is genetically modified for overexpression of at least one gene selected from the group consisting of:(a) a gene encoding an N-acetylglutamate kinase and N-acetyl-gamma-glutamyl-phosphate reductase;(b) a gene encoding a mitochondrial ornithine acetyltransferase;(c) a gene encoding an acetylornithine aminotransferase; and(d) a gene encoding an acetylglumate ...

Methods For Purification of Messenger RNA

The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, steps of forming an mRNA slurry, stirring the slurry, and vacuum or pressure filtering the slurry. 1. A method of purifying mRNA , comprising steps of:providing a solution comprising mRNA;adding one or more agents that promote precipitation of mRNA, thereby obtaining a slurry;stirring the slurry prior to and/or while providing pressure to the slurry and/or a vacuum to the slurry sufficient to direct the slurry's mother liquor through a filter, thereby obtaining a precipitate-containing composition; andwashing the precipitate-containing composition, thereby yielding a purified mRNA precipitate.2. A method for purifying at least about 1 , 2.5 , 5 , 10 , 25 , 50 , 100 or 1000 grams mRNA , comprising steps of:providing a solution comprising mRNA;adding one or more agents that promote precipitation of mRNA, thereby obtaining a slurry;stirring the slurry prior to and/or while providing pressure to the slurry and/or a vacuum to the slurry sufficient to direct the slurry's mother liquor through a filter, thereby obtaining a precipitate-containing composition; andwashing the precipitate-containing composition, thereby yielding a purified mRNA precipitate;wherein the total purified mRNA is recovered in an amount that results in a yield of at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95%; and/orthe total purified mRNA is substantially free of prematurely aborted RNA sequences and/or enzyme reagents used in in vitro synthesis.3. (canceled)4. The method of claim 1 , wherein at least the stirring step occurs in a stirred cell or in a Nutsche Filter.5. The method of claim 1 , wherein the step of the adding one or more agents that promote precipitation of mRNA occurs in a stirred cell or in the a Nutsche Filter.6. (canceled)7. The method of claim 1 , wherein the stirring occurs at a speed between about 50 RPM and about 500 RPM.8. (canceled)9. The method of ...

MESSENGER UNA MOLECULES AND USES THEREOF

This invention provides a range of translatable messenger UNA (mUNA) molecules. The mUNA molecules can be translated in vitro and in vivo to provide an active polypeptide or protein, or to provide an immunization agent or vaccine component. The mUNA molecules can be used as an active agent to express an active polypeptide or protein in cells or subjects. Among other things, the mUNA molecules are useful in methods for treating rare diseases. 1. A mUNA molecule , comprising one or more UNA monomers , and comprising nucleic acid monomers , wherein the mUNA molecule is translatable to express a polypeptide or protein.2. The molecule of claim 1 , wherein the molecule comprises from 200 to 12 claim 1 ,000 monomers.3. The molecule of claim 1 , wherein the molecule comprises from 200 to 4 claim 1 ,000 monomers.4. The molecule of claim 1 , wherein the molecule comprises from 1 to 8 claim 1 ,000 UNA monomers.5. The molecule of claim 1 , wherein the molecule comprises from 1 to 100 UNA monomers.6. The molecule of claim 1 , wherein the molecule comprises from 1 to 20 UNA monomers.7. The molecule of claim 1 , wherein the molecule comprises one or more modified nucleic acid nucleotides claim 1 , or one or more chemically-modified nucleic acid nucleotides.8. The molecule of claim 1 , wherein the molecule comprises a 5′ cap claim 1 , a 5′ untranslated region of monomers claim 1 , a coding region of monomers claim 1 , a 3′ untranslated region of monomers claim 1 , and a tail region of monomers.9. The molecule of claim 8 , wherein the molecule comprises a translation enhancer in a 5′ or 3′ untranslated region.10. The molecule of claim 1 , wherein the molecule is translatable in vivo.11. The molecule of claim 1 , wherein the molecule is translatable in vitro.12. The molecule of claim 1 , wherein the molecule is translatable in a mammalian cell.13. The molecule of claim 1 , wherein the molecule is translatable in a human in vivo.14. The molecule of claim 1 , wherein a translation ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. An adeno-associated virus (AAV)8 viral vector comprising an AAV8 capsid having packaged therein a heterologous gene operably linked to regulatory sequences which direct its expression , wherein the heterologous gene encodes factor IX , wherein the AAV8 capsid comprises a vp3 capsid protein having the sequence of aa 203 to 737 of SEQ ID NO: 2 , or a sequence which is at least 95% identical to said sequence of aa 203 to 737 of SEQ ID NO: 2.2. The vector according to claim 1 , further comprising one or more AAV inverted terminal repeat (ITR) sequence from an AAV heterologous to AAV8.3. The vector according to claim 2 , wherein the one or more AAV ITR is from AAV2.4. A composition comprising the vector according to and a pharmaceutically acceptable carrier.5. A host cell containing the vector according to in culture.6. A method for treating hemophilia B claim 1 , said method comprising the step of contacting a cell with a vector according to claim 1 , wherein said vector directs expression of factor IX.7. The method according to claim 6 , wherein said vector is delivered via intravenous administration.8. An adeno-associated virus (AAV)8 viral vector comprising an AAV8 capsid having packaged therein a heterologous gene operably linked to regulatory sequences which direct its expression claim 6 , wherein the heterologous gene encodes ornithine transcarbamylase (OTC) claim 6 , wherein the AAV8 capsid comprises a vp3 capsid protein having the sequence of aa 203 to 737 of SEQ ID NO: 2 claim 6 , or a sequence which is at least 95% identical to said sequence of aa 203 to 737 of SEQ ID NO: 2.9. The vector according to claim 8 , wherein said OTC is human OTC.10. The vector according to claim 8 , further comprising one or more AAV inverted terminal repeat (ITR) ...

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Non-viral delivery systems comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A composition comprising a non-viral carrier and a nucleic acid molecule comprising an engineered sequence encoding human ornithine transcarbamylase (hOTCase) and expression control sequences which direct expression of hOTCase in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over at least the mature hOTC sequence of SEQ ID NO: 1 , wherein said hOTC nucleic acid sequence is a DNA sequence of SEQ ID NO: 3.2. The composition according to claim 1 , wherein the expression control sequences further comprise a liver-specific promoter.3. The composition according to claim 2 , wherein the liver-specific promoter is selected from a thyroxin-binding globulin (TBG) promoter or a lymphocyte-specific protein 1 (LSP1) promoter.4. The composition according to claim 1 , wherein the expression cassette further comprises one or more of an intron claim 1 , a Kozak sequence claim 1 , a poly A claim 1 , and a posttranscriptional regulatory elements.5. The composition according to claim 1 , wherein nonviral carrier is a plasmid.6. The composition according to claim 1 , wherein the nucleic acid molecule is formulated in a moiety selected from the group consisting of micelles claim 1 , liposomes claim 1 , cationic lipid-nucleic acid compositions claim 1 , poly-glycan compositions claim 1 , block copolymers and other polymers claim 1 , lipid and/or cholesterol-based-nucleic acid conjugates.7. The composition according to claim 6 , wherein the moiety is a nanoparticle.8. A composition comprising an mRNA according to any one of SEQ ID NO: 10 claim 6 , ...

MICROORGANISMS HAVING PUTRESCINE PRODUCTIVITY AND PROCESS FOR PRODUCING PUTRESCINE USING THE SAME

The present invention relates to a recombinant microorganism capable of producing putrescine, in which the microorganism is modified to have enhanced NCgl2522 activity, thereby producing putrescine in a high yield, and a method for producing putrescine using the microorganism. 1. A method for producing putrescine , comprising:(i) culturing a recombinant microorganism having putrescine productivity, and(ii) recovering putrescine from the cultured recombinant microorganism or the cell culture,wherein the recombinant microorganism is modified to have enhanced activity of a protein having the amino acid sequence of SEQ ID NO: 21 or 23 as compared to the activity in the same microorganism without the modification.2. The method of claim 1 , wherein the recombinant microorganism further modified to have enhanced ornithine decarboxylase (ODC) activity as compared to the activity in the same microorganism without the modification.3. The method of claim 1 , wherein the recombinant microorganism further modified to have weakened activities of ornithine carbamoyltransferase (ArgF) and a protein involved in glutamate export claim 1 , as compared to the activity in the same microorganism without the modification.4. The method of claim 2 , wherein the ornithine decarboxylase (ODC) comprises the amino acid sequence of SEQ ID NO: 33.5. The method of claim 3 , wherein the ornithine carbamoyltransferase (ArgF) comprises the amino acid sequence of SEQ ID NO: 29 claim 3 , and the protein involved in glutamate export comprises the amino acid sequence of SEQ ID NO: 30.6. The method of claim 1 , wherein the recombinant microorganism is further modified to have enhanced activities of acetyl-gamma-glutamyl-phosphate reductase (ArgC) claim 1 , acetylglutamate synthase or ornithine acetyltransferase (ArgJ) claim 1 , acetylglutamate kinase (ArgB) claim 1 , and acetylornithine aminotransferase (ArgD) claim 1 , as compared to the activities in the same microorganism without the modification.7. ...

Methods For Purification of Messenger RNA

The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, a step of centrifuging an mRNA suspension in a centrifuge comprising a porous substrate at a speed sufficient to remove process contaminants and to precipitate purified mRNA composition onto the porous substrate. 1. A method for purifying mRNA , comprisingsteps of: providing a suspension comprisingprecipitated mRNA; andcentrifuging the suspension in a centrifuge comprising a porous substrate such that the precipitated mRNA is captured on the porous substrate, thereby purifying contaminants from the mRNA.37.-. (canceled)8. The method of claim 1 , further comprising a step of first producing the suspension of precipitated mRNA by providing a solution comprising mRNA and adding to the solution one or more agents that promote precipitation of mRNA.9. The method of claim 1 , wherein the suspension comprising precipitated mRNA comprises at least one filtration aid that is a dispersant.1012.-. (canceled)13. The method of claim 1 , wherein the speed of the centrifuging of the mRNA suspension is between about 2000 RPM and about 4000 RPM claim 1 , about 1000 RPM and about 5000 RPM claim 1 , about 2000 RPM and about 4000 RPM claim 1 , about 2000 RPM and about 3000 RPM claim 1 , or about 2500 RPM and about 3500 RPM.1417.-. (canceled)18. The method of claim 1 , further comprising adding one or more agents to the suspension that denature proteins and/or keep proteins soluble in an aqueous medium.1920.-. (canceled)21. The method of claim 1 , further comprising a step of washing the purified mRNA composition with a solvent.2226.-. (canceled)27. The method of claim 1 , further comprising a step of drying the captured mRNA.2830.-. (canceled)31. The method of claim 27 , wherein the dried purified mRNA is collected and stored at a temperature of or below about 0° C. for a time period of at least about one week to about two years claim 27 , and wherein the dried ...

Process of Preparing mRNA-Loaded Lipid Nanoparticles

The present invention provides an improved process for lipid nanoparticle formulation and mRNA encapsulation. In some embodiments, the present invention provides a process of encapsulating messenger RNA (mRNA) in lipid nanoparticles comprising a step of mixing a solution of pre-formed lipid nanoparticles and mRNA. 1. A process of encapsulating messenger RNA (mRNA) in lipid nanoparticles comprising: mixing a solution comprising pre-formed lipid nanoparticles and mRNA such that lipid nanoparticles encapsulating mRNA are formed.2. The process of claim 1 , wherein the solution comprising pre-formed lipid nanoparticles and mRNA comprises less than 10 mM citrate.3. The process of claim 1 , wherein the solution comprising pre-formed lipid nanoparticles and mRNA comprises less than 25% non-aqueous solvent.45-. (canceled)6. The process of claim 1 , comprising heating the lipid nanoparticles and mRNA to a temperature greater than ambient temperature before or after the mixing claim 1 , and wherein the temperature is or is greater than about 30° C. claim 1 , 37° C. claim 1 , 40° C. claim 1 , 45° C. claim 1 , 50° C. claim 1 , 55° C. claim 1 , 60° C. claim 1 , 65° C. claim 1 , or 70° C.78-. (canceled)9. The process of claim 1 , wherein the pre-formed lipid nanoparticles are formed by mixing lipids dissolved in ethanol with an aqueous solution.10. The process of claim 1 , wherein the lipids comprise one or more cationic lipids claim 1 , one or more helper lipids claim 1 , one or more cholesterol-based lipids and PEG lipids.11. The process of claim 10 , wherein the one or more cationic lipids are selected from the group consisting of cKK-E12 claim 10 , OF-02 claim 10 , C12-200 claim 10 , MC3 claim 10 , DLinDMA claim 10 , DLinkC2DMA claim 10 , ICE (Imidazol-based) claim 10 , HGT5000 claim 10 , HGT5001 claim 10 , HGT4003 claim 10 , DODAC claim 10 , DDAB claim 10 , DMRIE claim 10 , DOSPA claim 10 , DOGS claim 10 , DODAP claim 10 , DODMA and DMDMA claim 10 , DODAC claim 10 , DLenDMA ...

ARGININE SUPPLEMENTATION TO IMPROVE EFFICIENCY IN GAS FERMENTING ACETOGENS

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways. 1. A method for increasing the production of at least one ATP-intensive product , the method comprising;a. flowing a gaseous C1-containing substrate to a bioreactor containing a culture of a C1-fixing microorganism in a liquid nutrient media; andb. fermenting the culture to produce at least one product;wherein arginine is provided to the culture in excess of the cellular requirement of the culture; and wherein the C1-fixing microorganism comprises an arginine metabolism pathway.2. The method of wherein the arginine metabolism pathway comprises at least one of an arginine deaminase pathway and an arginine decarboxylase pathway claim 1 , wherein the arginine deaminase pathway comprises one or more enzymes selected from the group consisting of arginine deiminase (EC 3.5.3.6) claim 1 , ornithine carbomyltransferase (putrescine carbomyltransferase) (EC 2.1.3.3) and a carbamate kinase (EC 2.7.2.2) and the arginine decarboxylase pathway comprises one or more enzymes selected from the group consisting arginine decarboxylase (EC 4.1.1.19) claim 1 , putative arginine deaminase (EC 3.5.3.12) claim 1 , putrescine carbamoyl transferase (EC 2.1.3.6) and carbamate kinase (EC 2.7.2.2).3. The method of claim 1 , wherein arginine is provided to the culture in an amount ranging from the cellular requirement of the culture to about 1000 times the cellular requirement of the culture.4. The method of claim 1 , wherein arginine is provided to the culture in an amount ranging from 2 times the cellular requirement of the culture to 1000 times the cellular ...

Cleavable Lipids

Disclosed herein are novel compounds, pharmaceutical compositions comprising such compounds and related methods of their use. The compounds described herein are useful, e.g., as liposomal delivery vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and subsequent iransfection of said target cells, and in certain embodiments are characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids. 168.-. (canceled)69. A pharmaceutical composition comprising a lyophilized lipid nanoparticle , wherein the lipid nanoparticle comprises mRNA and one or more lyoprotectants selected from the group consisting of sucrose , trehalose , dextran and inulin.70. The pharmaceutical composition of claim 69 , comprising a lyoprotectant that is sucrose.71. The pharmaceutical composition of claim 69 , comprising a lyoprotectant that is trehalose.72. The pharmaceutical composition of claim 69 , comprising a lyoprotectant that is dextran.73. The pharmaceutical composition of claim 69 , comprising a lyoprotectant that is inulin.74. The pharmaceutical composition of claim 69 , comprising one or more cationic lipids.75. The pharmaceutical composition of claim 74 , comprising C12-200 claim 74 , DOTAP (1 claim 74 ,2-diolelyl-3-trimethylammonium propane) claim 74 , DODAP (1 claim 74 ,2-diolelyl-3-dimethylammonium propane) claim 74 , DOTMA (1 claim 74 ,2-di-O-octadecenyl-3-trimethylammonium propane) claim 74 , DlinDMA claim 74 , DLin-KC2-DMA claim 74 , HGT4003 claim 74 , or ICE ((3S claim 74 ,10R claim 74 ,13R claim 74 , 17R)-10 claim 74 ,13-dimethyl-17-((R)-6-methylheptan-2-yl)-2 claim 74 , 3 claim 74 , 4 claim 74 , 7 claim 74 , 8 claim 74 , 9 claim 74 , 10 claim 74 , 11 claim 74 , 12 claim 74 , 13 claim 74 , 14 claim 74 , 15 claim 74 , 16 claim 74 , 17-tetradecahydro-1H-cyclopenta[a]phenanthren-3-yl 3-(1H-imidazol-4-yl)propanoate).76. The pharmaceutical composition of claim 75 , wherein the cationic ...

Messenger rna therapy for the treatment of ornithine transcarbamylase deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase (OTC) deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase (OTC) protein at an effective dose and an administration interval such that at least one symptom or feature of the OTC deficiency is reduced in intensity, severity, or frequency or has delayed onset. In some embodiments, the mRNA is encapsulated in a liposome comprising one or more cationic lipids, one or more non-cationic lipids, one or more cholesterol-based lipids and one or more PEG-modified lipids.

Multimeric coding nucleic acid and uses thereof

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends.

Microorganisms for producing putrescine or ornithine and process for producing putrescine or ornithine using them

The present invention relates to a recombinant microorganism for producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same. Specifically, the present invention relates to a microorganism of the genus Corynebacterium capable of producing putrescine or ornithine, in which an activity of the transcriptional regulator of sugar metabolism (SugR) is weakened, an activity of the citrate synthase (GltA) is enhanced, or both are applied; and a method for producing putrescine or ornithine using the same.

COMPOSITIONS AND METHODS FOR TREATING ORNITHINE TRANSCARBAMYLASE DEFICIENCY

The present disclosure provides a modified human OTC protein having improved properties for the treatment of OTC deficiency in a patient. Preferably, the protein of the disclosure is produced from a codon optimized mRNA suitable for administration to a patient suffering from OTC deficiency wherein upon administration of the mRNA to the patient, the protein of the disclosure is expressed in the patient in therapeutically effective amounts to treat OTC deficiency. The present disclosure also provides codon optimized mRNA sequences encoding wild type human OTC comprising a 5′ UTR derived from a gene expressed by for use in treating OTC deficiency in a patient. 1. An ornithine transcarbamylase (OTC) protein comprising the amino acid sequence of SEQ ID NO:4 and having OTC enzymatic activity.2. A polynucleotide encoding the protein of .3. The polynucleotide of comprising an optimized coding region encoding SEQ ID NO: 4.4. The polynucleotide of claim 3 , wherein the polynucleotide is DNA.5. The polynucleotide of claim 3 , wherein the polynucleotide is an mRNA.6. (canceled)7. The polynucleotide of comprising a 3′ poly A tail having from about 60 sequential adenine nucleotides to about 125 sequential adenine nucleotides.8. (canceled)9. The polynucleotide of comprising a non-human 5′ untranslated region (5′UTR).10Arabidopsis thaliana.. The polynucleotide of claim 9 , wherein the 5′UTR is derived from a gene expressed by11. The polynucleotide of claim 10 , wherein the 5′ UTR comprises a sequence selected from SEQ ID NO: 6 claim 10 , SEQ ID NOS: 125-127 and SEQ ID NOS: 227-247.12. (canceled)13. The polynucleotide of comprising a 3′ untranslated region (3′ UTR) selected from the group consisting of: SEQ ID NOS: 16-22.14. (canceled)15. The polynucleotide of comprising a Kozak sequence of SEQ ID NO: 23 or a partial Kozak sequence of SEQ ID NO: 24.16. The polynucleotide of comprising a 5′ cap.18. The polynucleotide of claim 4 , wherein the optimized coding region comprises SEQ ID ...

Composition and Methods for Treatment of Ornithine Transcarbamylase Deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase protein at a low dose and at an administration interval such that at least one symptom or feature of the OTC deficiency is reduced. 1. A method of treating ornithine transcarbamylase (OTC) deficiency in a human , comprising administering to a human in need of treatment a pharmaceutical composition comprising an mRNA encoding an ornithine transcarbamylase protein formulated in a lipid nanoparticle ,wherein the mRNA encoding the ornithine transcarbamylase protein is administered at a therapeutic low dose of 0.5 mg/kg or less of mRNA at a dosing interval of once every two weeks or a longer dosing internal for a period sufficient to treat at least one symptom or reduce the level of a biomarker associated with ornithine transcarbamylase deficiency in the human relative to a control.2. The method of claim 1 , wherein the therapeutic low dose is 0.4 mg/kg or less claim 1 , or 0.3 mg/kg or less claim 1 , or 0.2 mg/kg or less claim 1 , or 0.15 mg/kg less claim 1 , or 0.10 mg/kg or less claim 1 , or 0.05 mg/kg or less claim 1 , or 0.01 mg/kg or less claim 1 , of mRNA encoding ornithine transcarbamylase protein.37-. (canceled)8. The method of claim 1 , wherein the longer dosing interval is once every three weeks or longer claim 1 , or once every 4 weeks or longer.9. The method of claim 1 , wherein the mRNA is codon optimized.10. The method of claim 1 , whereinthe symptom comprises hyperammonemia.11. The method of claim 1 , wherein the biomarker is selected from the group consisting of: high plasma ammonia level claim 1 , high tissue ammonia level claim 1 , urinary orotic acid claim 1 , citrulline claim 1 , serum glutamate claim 1 , brain myoinositol claim 1 , serum amino acids claim 1 , and combination thereof.1213-. (canceled)14. A method ...

ARGININE SUPPLEMENTATION TO IMPROVE EFFICIENCY IN GAS FERMENTING ACETOGENS

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways. 1. A method for increasing the production of at least one ATP-intensive product , the method comprising;a. flowing a gaseous C1-containing substrate to a bioreactor containing a culture of a C1-fixing microorganism in a liquid nutrient media; andb. fermenting the culture to produce at least one product;wherein arginine is provided to the culture in excess of the cellular requirement of the culture; and wherein the C1-fixing microorganism comprises an arginine metabolism pathway.2. The method of claim 1 , wherein arginine is provided to the culture in an amount ranging from 2 times the cellular requirement of the culture to 1000 times the cellular requirement of the culture.3. The method of claim 1 , wherein the cellular requirement of the culture is about 0.012 g of arginine per gram of cellular biomass.4. The method of claim 1 , wherein the doubling time of the culture is decreased by at least 10% when compared to a culture where arginine is not provided in excess of the cellular requirement of the culture.5. The method of claim 4 , wherein the doubling time of the culture is at decreased by at least 50% when compared to a culture where arginine is not provided in excess of the cellular requirement of the culture.6. The method of claim 1 , wherein selectivity to the ATP-intensive product is increased when compared to a culture where arginine is not provided in excess of the cellular requirement of the culture.7. The method of claim 1 , wherein productivity of the ATP-intensive product is at least 10% greater when compared to a ...

Stable Gene Transfer to Proliferating Cells

Provided herein are methods for facilitating or inducing stable transgene integration and expression in a proliferating cell, comprising administering to the cell (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences, which sequences are in turn flanked by AAV-derived inverted terminal repeat regions, and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell. Also provided are methods and transgene delivery systems for the treatment or prevention of diseases affecting, associated with or characterised by proliferating cells, including paediatric liver diseases, bone marrow diseases and cancer. 1. A method for stably integrating a transgene into the genome of a proliferating cell and/or inducing stable transgene expression in a proliferating cell , the method comprising administering to the cell: (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences , which sequences are in turn flanked by AAV-derived inverted terminal repeat regions; and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell.2. A method according to claim 1 , wherein the genomic integration of the transgene into the genome of the proliferating cell facilitates or induces the stable transgene expression.34-. (canceled)5. A method according to claim 1 , wherein the transgene and flanking transposon-derived inverted terminal repeat sequences form a transposon-transgene cassette claim 1 , optionally comprising one or more further sequences or genetic elements including claim 1 , for example claim 1 , a promoter claim 1 , enhancer claim 1 , post-regulatory element and/or ...

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification conferring stability to the RNA, as well as related methods, are disclosed. 1. A composition for delivery of mRNA for the expression of a protein in a target cell , comprising at least one mRNA molecule comprising a sequence encoding a protein of interest and a 3′ untranslated region (3′-UTR) comprising a sequence encoding a portion of human growth hormone (hGH).2. (canceled)3. The composition of claim 1 , wherein the mRNA molecule comprises more than one modification which confers stability to the mRNA molecule.4. The composition of claim 1 , wherein the mRNA molecule comprises a modification of the 5′ untranslated region.5. The composition of claim 4 , wherein said modification comprises a partial sequence of a CMV immediate-early 1 (IE 1) gene.6. The composition of claim 5 , wherein said partial sequence of the CMV immediate-early 1 (IEI) gene comprises SEQ ID NO: 2 or SEQ ID NO: 1.7. The composition of claim 1 , wherein the mRNA comprises the inclusion of a poly A tail.8. The composition of claim 1 , wherein the mRNA comprises the inclusion of a Cap 1 structure.910-. (canceled)11. The composition of claim 1 , wherein the sequence encoding a portion of human growth hormone (hGH) comprises SEQ ID NO: 3.12. (canceled)13. The composition of claim 1 , wherein the mRNA encodes ornithine carbamoyl transferase.14. The composition of claim 1 , wherein the mRNA encodes alpha galactosidase.15. The composition of claim 1 , wherein the mRNA encodes erythropoietin.16. A method of treating a subject deficient in a protein claim 1 , comprising administering a composition comprising an mRNA and a transfer vehicle claim 1 , wherein the mRNA encodes a protein corresponding to the protein which is deficient in the subject claim 1 , and wherein the mRNA comprises comprises a 3′ untranslated region (3′-UTR) comprising a sequence ...

Cleavable Lipids

Disclosed herein are novel compounds, pharmaceutical compositions comprising such compounds and related methods of their use. The compounds described herein are useful, e.g., as liposomal delivery vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and subsequent iransfection of said target cells, and in certain embodiments are characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids.

METHODS FOR PURIFICATION OF MESSENGER RNA

The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, a step of centrifuging an mRNA suspension in a centrifuge comprising a porous substrate at a speed sufficient to remove process contaminants and to precipitate purified mRNA composition onto the porous substrate. 1. A method for purifying mRNA , comprising steps of:providing a suspension comprising precipitated mRNA; andcentrifuging the suspension in a centrifuge comprising a porous substrate such that the precipitated mRNA is captured on the porous substrate, thereby purifying contaminants from the mRNA.2. The method of claim 1 , comprisingpurifying at least about 10 grams, 25 grams, 50 grams, 100 grams, or 1 kilogram mRNA,wherein the total purified mRNA is recovered in an amount that results in a yield of at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95%; and/orthe total purified mRNA is substantially free of prematurely aborted RNA sequences and/or enzyme reagents used in in vitro synthesis.37.-. (canceled)8. The method of claim 1 , further comprising a step of first producing the suspension of precipitated mRNA by providing a solution comprising mRNA and adding to the solution one or more agents that promote precipitation of mRNA.9. The method of claim 1 , wherein the suspension comprising precipitated mRNA comprises at least one filtration aid that is a dispersant.1012.-. (canceled)13. The method of claim 1 , wherein the speed of the centrifuging of the mRNA suspension is between about 2000 RPM and about 4000 RPM claim 1 , about 1000 RPM and about 5000 RPM claim 1 , about 2000 RPM and about 4000 RPM claim 1 , about 2000 RPM and about 3000 RPM claim 1 , or about 2500 RPM and about 3500 RPM.1417.-. (canceled)18. The method of claim 1 , further comprising adding one or more agents to the suspension that denature proteins and/or keep proteins soluble in an aqueous medium.1920.-. (canceled)21. The method of claim 1 , further comprising a ...

Methods For Purification of Messenger RNA

The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, steps of forming an mRNA slurry, stirring the slurry, and vacuum or pressure filtering the slurry. 1. A method of purifying mRNA , comprising steps of:providing a solution comprising mRNA;adding one or more agents that promote precipitation of mRNA, thereby obtaining a slurry;stirring the slurry prior to and/or while providing pressure to the slurry and/or a vacuum to the slurry sufficient to direct the slurry's mother liquor through a filter, thereby obtaining a precipitate-containing composition; andwashing the precipitate-containing composition, thereby yielding a purified mRNA precipitate.2. The method of claim 1 , comprising purifying at least about 1 claim 1 , 2.5 claim 1 , 5 claim 1 , or 10 grams mRNA or at least about 25 claim 1 , 50 claim 1 , 100 claim 1 , or 1000 grams mRNA claim 1 ,wherein the total purified mRNA is recovered in an amount that results in a yield of at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, or 95%; and/or the total purified mRNA is substantially free of prematurely aborted RNA sequences and/or enzyme reagents used in in vitro synthesis.3. (canceled)4. The method of claim 1 , wherein at least the stirring step occurs in a stirred cell or in a Nutsche Filter.5. The method of claim 1 , wherein the step of the adding one or more agents that promote precipitation of mRNA occurs in the stirred cell or in the Nutsche Filter.6. The method of claim 1 , wherein the step of adding one or more agents that promote precipitation of mRNA does not occur in the stirred cell or in the Nutsche Filter.7. The method of claim 1 , whereinthe stirring occurs at a speed between about 50 RPM and about 500 RPM;the pressure is between about 5 PSI and about 100 PSI; ora washing step is diafiltering, ultrafiltering, or dialyzing.814.-. (canceled)15. The method of claim 1 , further comprising adding one or more agents that denatures proteins ...

MULTIMERIC CODING NUCLEIC ACID AND USES THEREOF

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends. 142-. (canceled)43. A method of delivering a multimeric coding nucleic acid (MCNA) for in vivo protein production , comprising administering to a subject in need of delivery a MCNA , wherein the MCNA comprises two messenger RNAs (mRNAs) linked at 3′ ends , via stable linkage , such that the multimeric coding nucleic acid has two 5′ ends , and wherein the stable linkage is an oligonucleotide bridge comprising an internal 3′-to-3′ inverted phosphodiester linkage.4447-. (canceled)48. The method of claim 43 , wherein each of the mRNAs encodes a protein of interest.49. The method of claim 48 , wherein each of the mRNAs encodes a same protein.50. The method of claim 48 , wherein each of the mRNAs encodes a distinct protein.51. The method of claim 43 , wherein the mRNAs comprise a 3′ UTR.52. The method of claim 51 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.53. The method of claim 43 , wherein the oligonucleotide bridge comprises nucleosides selected from the group consisting of 2′-OMe-A claim 43 , 2′-OMe-G claim 43 , 2′-OMe-C claim 43 , 2′-OMe-U claim 43 , 2′-F-A claim 43 , 2′-F-G claim 43 , 2′-F-C claim 43 , 2′-F-U claim 43 , LNA-A claim 43 , LNA-G claim 43 , LNA-C claim 43 , LNA-U claim 43 , N6-methyl-adenosine claim 43 , 2-thiouridine (2sU) claim 43 , 5-methyl-cytidine (5mC) claim 43 , pseudouridine (ψU) claim 43 , and 1-methyl-pseudouridine.54. The method of claim 43 , wherein the mRNAs comprise one or more modified nucleosides.55. The method of claim 54 , wherein the modified nucleosides are selected from the group consisting of 2′-OMe-A claim 54 , 2′-OMe-G claim 54 , 2′- ...

ENZYMATIC SYSTEMS FOR CARBON FIXATION AND METHODS OF GENERATING SAME

A system for carbon fixation is provided. The system comprises enzymes which catalyze reactions of a carbon fixation pathway, wherein at least one of the reactions of the carbon fixation pathway is a carboxylation reaction, wherein products of the reactions of the carbon fixation pathway comprise oxaloacetate and malonyl-CoA, wherein an enzyme which performs the carboxylation reaction is selected from the group consisting of phophoenolpyruvate (PEP) carboxlase, pyruvate carboxylase and acetyl-CoA carboxylase and wherein an export product of the carbon fixation pathway is glyoxylate. Additional carbon fixation pathways are also provided and methods of generating same. 1. A system for carbon fixation , comprising an electron donor and enzymes which catalyze reactions of a carbon fixation pathway , wherein all the carboxylation reactions of the carbon fixation pathway utilize:(i) phophoenolpyruvate (PEP) carboxylase and acetyl-CoA carboxylase; or(ii) phophoenolpyruvate (PEP) carboxylase; or(iii) pyruvate carboxylase and acetyl-CoA carboxylase; or(iv) pyruvate carboxylasewherein products of said reactions of the carbon fixation pathway comprise oxaloacetate and malonyl-CoA, and wherein an additional product of the carbon fixation pathway is glyoxylate, wherein an in-organic carbon is introduced into a substrate to become a carboxylic acid group during said carboylation reactions.2. The system of claim 1 , wherein said glyoxylate is an export product.3. The system of claim 1 , wherein an export product of the carbon fixation pathway is pyruvate.4. The system of claim 1 , wherein the enzymes of the carbon fixation pathway generate more than 0.3 μmol glyceraldehyde-3-phosphate/min/mg.5. The system of claim 1 , wherein said enzyme which performs said carboxylation enzyme is PEP carboxylase.6. The system of claim 1 , wherein at least two of said reactions of the carbon fixation pathway are carboxylation reactions.7. The system of claim 1 , wherein one of said reactions of ...

METHODS OF DETERMINING DOSING OF A THERAPEUTIC AGENT BASED ON MEASURED LEVELS OF A METABOLITE

The invention provides methods of determining a therapeutically effective dose of an agent that targets a metabolic pathway based on measured levels of a metabolite in the pathway. The methods, which may include providing the agent in a therapeutically effective dose, are useful for treating disorders, such as cancer, in a subject. The invention also provides methods for assessing the impact of a therapeutic agent on a tumor in a subject by monitoring, in real time, metabolism of a molecule in the tumor, oxygenation of the tumor, or both. The invention further provides devices that determine a therapeutically effective dose of an agent that targets a metabolic pathway based on measured levels of a metabolite in the pathway and notify a subject to administer the dose. 1. A method for determining a therapeutically effective dose of an agent to treat a disorder , the method comprising:receiving information regarding a measured level of a metabolite in a metabolic pathway in a sample from a subject having a disorder;comparing the received information to a reference that provides an association of a measured level of the metabolite with a recommended dosage adjustment of an agent; anddetermining, based on the comparing step, a dosage of the agent that results in the level of the metabolite being raised or maintained above a threshold level, the threshold level being indicative that a sufficient amount of the agent is present in the subject to sufficiently alter the metabolic pathway to ameliorate, reduce, or eliminate at least one sign or symptom of the disorder.2. The method of claim 1 , wherein the recommended dosage adjustment is at least one selected from the group consisting of: increase the dosage by a certain value claim 1 , decrease the dosage by a certain value claim 1 , and make no adjustment to the dosage.3. The method of claim 1 , wherein the agent inhibits an enzyme in the metabolic pathway.4. The method of claim 3 , wherein the metabolite is a substrate of ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. (canceled)2. A host cell comprising a recombinant nucleic acid molecule:(a) encoding an AAV8 vp1 capsid protein having a sequence comprising amino acids 1 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2121 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2121 to 4334 of SEQ ID NO: 1, a functional rep gene, a minigene comprising AAV inverted terminal repeats (ITRs) and a transgene, and sufficient helper functions to permit packaging of the minigene into the AAV capsid, wherein the transgene encodes factor IX.3. The host cell according to claim 1 , wherein said recombinant nucleic acid molecule comprises a sequence selected from:vp1, nt 2121 to 4334;vp2, nt 2532 to 4334; orvp3, nt 2730 to 4334 of SEQ ID NO: 1.4. The host cell according to claim 1 , wherein said recombinant nucleic acid molecule is a plasmid.5. The host cell according to claim 1 , which further comprises a nucleic acid sequence(a) encoding an AAV8 vp2 capsid protein having a sequence comprising amino acids 138 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2532 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2532 to 4334 of SEQ ID NO: 1.6. The host cell according to claim 1 , which further comprises a nucleic acid sequence(a) encoding an AAV8 vp3 capsid protein having a sequence comprising amino acids 204 to 738 of SEQ ID NO: 2; or(b) comprising nucleotides 2730 to 4334 of SEQ ID NO: 1, or a nucleotide sequence at least 99% identical to nucleotides 2730 to 4334 of SEQ ID NO: 1.7. An AAV8 vector comprising an AAV8 capsid claim 1 , said capsid having packaged therein a nucleic acid molecule comprising AAV inverted terminal repeat sequences and a gene encoding factor IX claim 1 , which gene is operably ...

ADENO-ASSOCIATED VIRUS (AAV) SEROTYPE 8 SEQUENCES, VECTORS CONTAINING SAME, AND USES THEREFOR

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles. 1. An adeno-associated virus (AAV)8 viral vector comprising an AAV8 capsid having packaged therein a heterologous gene operably linked to regulatory sequences which direct its expression, wherein the heterologous gene encodes factor IX. This application is a continuation of U.S. patent application Ser. No. 15/084,615, filed Mar. 30, 2016, which is a divisional of U.S. patent application Ser. No. 14/598,462, filed Jan. 16, 2015, now U.S. Pat. No. 9,493,788, issued Nov. 15, 2016, which is a divisional of U.S. patent application Ser. No. 11/981,022, filed Oct. 31, 2007, now U.S. Pat. No. 8,962,330, issued Feb. 24, 2015, which is a continuation of U.S. patent application Ser. No. 11/899,500, filed Sep. 6, 2007, now U.S. Pat. No. 7,790,449, issued Sep. 7, 2010, which is a continuation of U.S. patent application Ser. No. 10/423,704, filed Apr. 25, 2003, now U.S. Pat. No. 7,282,199, issued Oct. 16, 2007, which is a continuation-in-part of International Patent Application No. PCT/US02/33630, filed Nov. 12, 2002, which claims the benefit under 35 USC 119(e) of U.S. Provisional Patent Application No. 60/386,122, filed Jun. 5, 2002, U.S. Provisional Patent Application No. 60/377,133, filed May 1, 2002, and U.S. Provisional Patent Application No. 60/341,151, filed Dec. 17, 2001, which applications are incorporated herein by reference in their entireties.This invention was made with government support under Grant No. P30 DK 47757-09 awarded by the National Institute of Diabetes and Digestive and Kidney Diseases, and Grant No. P01 HL 59407-03 awarded by the National Heart, Lung, and Blood Institute. The government has certain rights in the invention.The Sequence Listing material filed in electronic form herewith is hereby incorporated by reference. This file is labeled ...

SUBCUTANEOUS DELIVERY OF MESSENGER RNA

The present invention provides, among other things, methods of formulating nucleic acid-containing nanoparticles with an enzyme to afford efficient delivery of payload to a cell or tissue of interest via subcutaneous administration. In some embodiments, the present invention provides a process in which mRNA-loaded lipid nanoparticles are co-mixed with various amounts of hyaluronidase and administered via subcutaneous administration. The resulting payload can be efficiently delivered to the liver and other organs or tissues of a treated subject. 1. A method of treating ornithine transcarbamylase (OTC deficiency) comprising administering via subcutaneous injection to a subject in need of treatmenta) an mRNA encoding an ornithine transcarbamylase (OTC) protein, andb) a hyaluronidase enzyme.2. The method of claim 1 , wherein the subcutaneous injection results in delivery of mRNA in the liver.3. The method of or claim 1 , wherein the subcutaneous injection results in expression of the OTC protein in the liver of the subject.4. The method of any one of the preceding claims claim 1 , wherein the subcutaneous injection results in expression of the OTC protein in the serum of the subject.5. The method of or claim 1 , wherein the OTC protein is detectable after at least 24 hours claim 1 , 2 days claim 1 , 3 days claim 1 , 4 days claim 1 , 5 days claim 1 , 6 days claim 1 , 1 week claim 1 , 2 weeks claim 1 , 3 weeks claim 1 , 4 weeks claim 1 , or 1 month post-injection.6. The method of claim 5 , wherein the OTC protein is detected by a functional assay.7. The method of any one of the preceding claims claim 5 , wherein the hyaluronidase enzyme is administered at a dose amount of less than 50 claim 5 ,000 U.8. The method of claim 7 , wherein the hyaluronidase enzyme is administered at a dose amount of less than 40 claim 7 ,000 U claim 7 , Less than 30 claim 7 ,000 U claim 7 , less than 20 claim 7 ,000 U claim 7 , less than 10 claim 7 ,000 U claim 7 , less than 9000 U claim 7 , ...

POLYNUCLEOTIDES ENCODING ORNITHINE TRANSCARBAMYLASE FOR THE TREATMENT OF UREA CYCLE DISORDERS

This disclosure relates mRNA therapy for the treatment of ornithine transcarbamylase deficiency (OTCD). mRNAs for use in the invention, when administered in vivo, encode human ornithine transcarbamylase (OTC), isoforms thereof, functional fragments thereof, and fusion proteins comprising OTC. mRNAs of the invention are preferably encapsulated in lipid nanoparticles (LNPs) to effect efficient delivery to cells and/or tissues in subjects, when administered thereto. mRNA therapies of the invention increase and/or restore deficient levels of OTC expression and/or activity in subjects. mRNA therapies of the invention further decrease levels of toxic ammonia associated with deficient OTC activity in subjects. 1. A pharmaceutical composition comprising an mRNA comprising an open reading frame (ORF) encoding an ornithine transcarbamylase (OTC) polypeptide , wherein the composition when administered as a single intravenous dose to a human subject in need thereof is sufficient to:(i) increase the level of OTC activity in liver tissue to within at least 2%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 100% of normal OTC activity level for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at least 4 weeks post-administration;(ii) increase the level of OTC activity in liver tissue at least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 10-fold, at least 20-fold, or at least 50-fold compared to the subject's baseline OTC activity level or a reference OTC activity level in a human subject having ornithine transcarbamylase deficiency (OTCD) for at least 12 hours, at least 24 hours, at least 48 hours, at least 72 hours, at least 96 hours, at least 120 hours, at least 1 week, at least 2 weeks, at least 3 weeks, or at ...

MULTIMERIC CODING NUCLEIC ACID AND USES THEREOF

The present invention provides, among other things, multimeric coding nucleic acids that exhibit superior stability for in vivo and in vitro use. In some embodiments, a multimeric coding nucleic acid (MCNA) comprises two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends. 1. A multimeric coding nucleic acid (MCNA) comprising two or more encoding polynucleotides linked via 3′ ends such that the multimeric coding nucleic acid compound comprises two or more 5′ ends.23-. (canceled)4. The MCNA of claim 1 , wherein each of the two or more encoding polynucleotides is a synthetic polydeoxyribonucleotide or a polyribonucleotide.5. The MCNA of claim 1 , wherein each of the two or more encoding polynucleotides encodes a protein of interest.6. The MCNA of claim 5 , wherein each of the two or more encoding polynucleotides encodes a same protein.7. The MCNA of claim 5 , wherein each of the two or more encoding polynucleotides encodes a distinct protein.8. The MCNA of claim 1 , wherein the compound comprises three or more encoding polynucleotides.911-. (canceled)1211. The MCNA of claim claim 1 , wherein the one or more of the encoding polynucleotides comprise a 3′ UTR.13. (canceled)14. The MCNA of claim 12 , wherein the 3′ UTR comprises a plurality of multi-A segments with spacers in between.1525-. (canceled)26. The MCNA of claim 1 , wherein the 3′ ends of the two or more encoding polynucleotides are linked via an oligonucleotide bridge comprising a 3′-3′ inverted phosphodiester linkage.27. The MCNA of claim 26 , wherein the nucleotides comprising the oligonucleotide bridge are selected from the group consisting of 2′-OMe-A claim 26 , 2′-OMe-G claim 26 , 2′-OMe-C claim 26 , 2′-OMe-U claim 26 , 2′-F-A claim 26 , 2′-F-G claim 26 , 2′-F-C claim 26 , 2′-F-U claim 26 , LNA-A claim 26 , LNA-G claim 26 , LNA-C claim 26 , LNA-U claim 26 , N6-methyl-adenosine claim 26 , 2-thiouridine (2sU) claim 26 , 5- ...

ORNITHINE TRANSCARBAMYLASE CODING POLYRIBONUCLEOTIDES AND FORMULATIONS THEREOF

In certain aspects, the disclosure relates to compositions comprising modified Ornithine transcarbamylase (OTC) polyribonucleotides and methods of use. 1. A modified polyribonucleotide comprising a primary sequence which that is at least 95% identical to SEQ ID NO: 4 and which encodes an ornithine transcarbamylase (OTC) protein , wherein the modified polyribonucleotide contains a combination of unmodified and modified ribonucleotides , wherein 30-50% of the uridines are analogs of uridine and 5-30% of the cytidines are analogs of cytidine.2. (canceled)3. (canceled)4. The modified polyribonucleotide of claim 1 , wherein the modified polyribonucleotide contains a combination of unmodified and modified ribonucleotides claim 1 , wherein 30-45% of the uridines are analogs of uridine and 5-20% of the cytidines are analogs of cytidine; orwherein the modified polyribonucleotide is made using an input mixture of ribonucleotides, wherein 30-45% of the uridines are analogs of uridine and 5-20% of the cytidines are analogs of cytidine.5. (canceled)6. A modified polyribonucleotide comprising a primary sequence at least 99% identical to a sequence selected from the group consisting of SEQ ID NOs: 21-27 claim 1 , wherein the modified polyribonucleotide contains a combination of unmodified and modified ribonucleotides claim 1 , wherein 5-50% of the uridines are analogs of uridine and 5-50% of the cytidines are analogs of cytidine.7. The modified polyribonucleotide of claim 6 , wherein 25-50% of uridines are analogs of uridine and 5-30% of cytidines are analogs of cytidine.8. (canceled)9. (canceled)10. (canceled)11. (canceled)12. The modified polyribonucleotide of claim 1 , wherein the modified polyribonucleotide is codon-optimized for expression in mammalian cells.13. (canceled)14. The modified polyribonucleotide of claim 1 , wherein the modified polyribonucleotide further comprises a 3′ UTR claim 1 , a 5′ UTR claim 1 , or a 3′ UTR and a 5′ UTR claim 1 , and wherein the UTR(s) aid( ...

FUSION PROTEINS

Provided are fusion target-binding proteins comprising a target binding moiety, an intracellular signalling region and a domain that promotes synthesis of arginine or an arginine precursor. The domain may be an enzyme domain such as an argininosuccinate synthase (ASS-1) enzyme domain, or an ornithine transcarbamylase (OTC) enzyme domain. Also provided are cells comprising such a fusion target-binding protein (for example cells that express the fusion target-binding protein), and nucleic acids encoding such fusion target-binding proteins. The invention also provides fusion target-binding proteins comprising a target binding moiety, an intracellular signalling region and a domain that promotes synthesis of tryptophan or a tryptophan precursor. Pharmaceutical compositions, medical uses, and methods of treatment, all using the fusion target-binding proteins, cells, or nucleic acids are disclosed. The proteins, cells, nucleic acids and pharmaceutical compositions may be used in the prevention and/or treatment of cancer, such as neuroblastoma or acute myeloid leukaemia. 1. A fusion target-binding protein comprising a target binding moiety , an intracellular signalling region and a domain that promotes synthesis of arginine or an arginine precursor.2. The fusion target-binding protein according to claim 1 , wherein the domain that promotes synthesis of arginine or an arginine precursor comprises an enzyme domain.3. The fusion target-binding protein according to claim 2 , wherein the enzyme domain is selected from the group consisting of: an ASS-1 domain; an OTC domain; an ASL domain; an OCD1 domain; an ArgG domain; an ArgH domain; and an ArgF domain.4. The fusion target-binding protein according to claim 3 , wherein the ASS-1 domain comprises the amino acid sequence of SEQ ID NO. 1.5. The fusion target-binding protein according to claim 3 , wherein the OTC domain comprises the amino acid sequence of SEQ ID NO. 2 claim 3 , the ASL domain comprises the amino acid sequence ...

POLYNUCLEOTIDES AND VECTORS FOR THE EXPRESSION OF TRANSGENES

Polynucleotides and vectors can be used for the expression of a transgene in cells, such as liver cells. The expression of the transgene from the polynucleotides and vectors can be useful in gene therapy. Various methods can be used for expressing the transgene from the polynucleotides and vectors in liver cells. 1. A polynucleotide , comprising , from 5′ to 3′ , a human ornithine transcarbamylase (hOTC) enhancer , a liver-specific promoter and a transgene , wherein the hOTC enhancer is operably linked to the liver-specific promoter and the liver-specific promoter is operably linked to the transgene.2. The polynucleotide of claim 1 , further comprising an intron between the liver-specific promoter and the transgene.3. The polynucleotide of claim 2 , wherein the intron is a SV40 intron or a beta-globin intron.4. The polynucleotide of any one of - claim 2 , further comprising a polyadenylation signal sequence 3′ of the transgene.5. The polynucleotide of claim 4 , wherein the polyadenylation signal sequence is a BGH-poly(A) signal.6. The polynucleotide of any one of - claim 4 , further comprising a Kozac sequence between the liver-specific promoter and the transgene.7. The polynucleotide of any one of - claim 4 , comprising claim 4 , from 5′ to 3′ claim 4 , a hOTC enhancer claim 4 , a liver-specific promoter claim 4 , an intron claim 4 , a Kozac sequence claim 4 , a transgene claim 4 , and a polyadenylation signal.8. The polynucleotide of any one of - claim 4 , comprising two or more hOTC enhancers operably linked to the liver-specific promoter.9. The polynucleotide of any one of - claim 4 , wherein the liver-specific promoter is a hOTC promoter or a human alpha 1-antitrypsin (hAAT) promoter.10. The polynucleotide of claim 9 , comprising claim 9 , from 5′ to 3′ claim 9 , a hOTC enhancer claim 9 , a hOTC promoter claim 9 , a SV40 intron claim 9 , a Kozac sequence claim 9 , a transgene claim 9 , and a BGH-poly(A) signal.11. The polynucleotide of claim 9 , comprising ...

COMPOSITIONS USEFUL IN TREATMENT OF ORNITHINE TRANSCARBAMYLASE (OTC) DEFICIENCY

Viral vectors comprising engineered hOTC DNA and RNA sequences are provided which when delivered to a subject in need thereof are useful for treating hyperammonemia, ornithine transcarbamylase deficiency and symptoms associated therewith. Also provided are methods of using hOTC for treatment of liver fibrosis and/or cirrhosis in OTCD patients by administering hOTC. 1. A composition comprising a nucleic acid sequence encoding human ornithine transcarbamylase (hOTCase) and expression control sequences which direct expression of hOTC in a liver cell , wherein the hOTC nucleic acid sequence is less than 80% identical to the wild-type hOTC sequence over the mature sequence or full length hOTC of SEQ ID NO:1 , and expresses a functional hOTCase , wherein said hOTC nucleic acid sequence is selected from the nucleic acid sequence comprising SEQ ID NO: 5 or a nucleic acid sequence at least about 96 to about 99% identical thereto or a nucleic acid sequence selected from SEQ ID NO: 9 , or a nucleic acid sequence at least about 96 to about 99% identical thereto.2. The recombinant viral vector according to claim 1 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 4.3. The recombinant viral vector according to claim 1 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 3.4. The recombinant viral vector according to claim 1 , wherein the hOTC nucleic acid sequence has the sequence of SEQ ID NO: 8.5. The recombinant viral vector according to claim 1 , wherein the hOTC is a chimeric OTC comprises a heterologous transit sequence substituted for the native transit sequence of SEQ ID NO: 5 or 9.6. The recombinant viral vector according to claim 1 , wherein the viral vector is selected from an adeno-associated virus (AAV) vector claim 1 , an adenoviral vector claim 1 , and a lentiviral vector.7. The recombinant viral vector according to claim 1 , wherein the expression control sequences further comprise a liver-specific promoter.8. The ...

Production of odd chain fatty acid derivatives in recombinant microbial cells

Recombinant microbial cells are provided which have been engineered to produce fatty acid derivatives having linear chains containing an odd number of carbon atoms by the fatty acid biosynthetic pathway. Also provided are methods of making odd chain fatty acid derivatives using the recombinant microbial cells, and compositions comprising odd chain fatty acid derivatives produced by such methods.

Microorganisms for the production of putrescine or ornithine and procedure for the production of putrescine or ornithine using the same

<p>SE DESVELA UN MICROORGANISMO MODIFICADO QUE PRODUCE PUTRESCINA U ORNITINA, Y UN PROCEDIMIENTO PARA LA PRODUCCIÓN DE PUTRESCINA U ORNITINA USANDO EL MISMO.</p> <p> A MODIFIED MICROORGANISM THAT PRODUCES PUTRESCINA OR ORNITINE, AND A PROCEDURE FOR THE PRODUCTION OF PUTRESCINA OR ORNITINE USING THE SAME IS DEVELOPED. </p>

Recombinant microorganism producing itaconic acid and production method of itaconic acid using the same

The present invention relates to a recombinant microorganism for itaconic acid. More particularly, the present invention relates to a recombinant microorganism for producing itaconic acid, with improved productivity of itaconic acid through regulation of gene expression involved in acetic acid metabolism, and a production method of itaconic acid using the same. The recombinant microorganism for producing itaconic acid according to the present invention can enhance economic feasibility of itaconic acid by remarkably increasing the production amount of itaconic acid by regulating gene expression involved in acetic acid metabolism, and thus can be used in various fields such as food additives, latex, and synthetic resins where itaconic acid is used.

Arginine supplementation to improve efficiency in gas fermenting acetogens

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways.

Corynebacterium glutamicum variety producing l-arginine and method for fabricating the same

An L-arginine producing microorganism is provided to over-express an argF2 gene, thereby producing an L-arginine with high yield and being usefully used for human medicinal and pharmaceutical industries. An L-arginine producing microorganism is characterized in that it is transformed by a polynucleotide encoding an argF(NCBI access no. Ncg10990) polypeptide. In the microorganism, the argF2 polypeptide has an amino acid sequence described as SEQ ID : NO. 1, the polynucleotide is described as SEQ ID : NO. 2 and the microorganism is Corynebacterium sp. and is deposited as a deposition no. KCCM10819P or KCCM10821P. The L-arginine producing microorganism is cultured in order to produce L-arginine.

Putrescine producing microorganisms, and the putrescine production method by using these microorganisms

FIELD: chemistry.SUBSTANCE: group of inventions relates to the putrescine producing microorganism, and to the use of said microorganism putrescine production method. In the proposed microorganism, the protein activity is increased having the amino acid sequence, as presented in the SEQ ID NO: 21 or 23, compared to the said protein activity in the wild-type microorganism. Putrescine production method includes the said microorganism culturing with the cells culture production, and the putrescine isolation from the cultured microorganism or cells culture.EFFECT: group of inventions provides putrescine production in high yield.10 cl, 11 tbl, 6 ex, 2 dwg РОССИЙСКАЯ ФЕДЕРАЦИЯ ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) (19) RU (11) (13) 2 665 825 C2 (51) МПК C12N 1/21 (2006.01) C12N 15/52 (2006.01) C12N 9/02 (2006.01) C12N 9/10 (2006.01) C12N 9/12 (2006.01) C12N 9/88 (2006.01) C12P 13/00 (2006.01) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/52 (2006.01); C12N 9/0008 (2006.01); C12N 9/1018 (2006.01); C12N 9/1029 (2006.01); C12N 9/1096 (2006.01); C12N 9/1217 (2006.01); C12N 9/88 (2006.01); C12P 13/001 (2006.01); C12Y 102/01038 (2006.01); C12Y 201/03003 (2006.01); C12Y 203/01035 (2006.01); C12Y 206/01011 (2006.01); C12Y 207/02008 (2006.01); C12Y 401/01007 (2006.01) 2015142261, 25.02.2014 (24) Дата начала отсчета срока действия патента: 25.02.2014 04.09.2018 Приоритет(ы): (30) Конвенционный приоритет: 20.03.2013 KR 10-2013-0030020; 14.02.2014 KR 10-2014-0017243 (43) Дата публикации заявки: 26.04.2017 Бюл. № 12 (56) Список документов, цитированных в отчете о поиске: WO 2012077995 A2, 14.06.2012. RU (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.10.2015 2433180 C2, 10.11.2011. SCHNEIDER J. ET AL. Improving putrescine production by Corynebacterium glutamicum by fine-tuning ornithine transcarbamoylase activity using a plasmid addiction system // Appl Microbiol Biotechnol (2012) 95:169-178. ZAHOOR A. ET AL. Metabolic engineering of ...

Microorganisms for producing ornithine and method of producing ornithine using said microorganisms

FIELD: biotechnology. SUBSTANCE: present invention relates to biotechnology and represents a modified microorganism of the genus Corynebacterium , which produces ornithine, where the activity of N-acetylglutamate synthase from E. coli and acetylornithine deacetylase from E. coli are introduced into said microorganism of the genus Corynebacterium , having acetylglutamate synthase or ornithine acetyltransferase activity (ArgJ). EFFECT: invention increases ornithine production. 12 cl, 2 dwg, 11 tbl, 7 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 721 852 C1 (51) МПК C12N 15/77 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/52 (2020.02); C12N 9/1029 (2020.02); C12N 9/80 (2020.02); C12N 9/88 (2020.02); C12P 13/001 (2020.02) (21)(22) Заявка: 2019126098, 19.07.2016 19.07.2016 Дата регистрации: 25.05.2020 20.07.2015 KR 10-2015-0102624 Номер и дата приоритета первоначальной заявки, из которой данная заявка выделена: 2018104512 20.07.2015 Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" (54) Микроорганизмы для получения орнитина и способ получения орнитина с использованием указанных микроорганизмов (57) Реферат: Настоящее изобретение относится к указанный микроорганизм рода Corynebacterium, биотехнологии и представляет собой обладающий активностью модифицированный микроорганизм рода ацетилглутаматсинтазы или Corynebacterium, продуцирующий орнитин, где орнитинацетилтрансферазы (ArgJ). Изобретение активности N-ацетилглутаматсинтазы из E. coli и позволяет повысить продукции орнитина. 2 н. и ацетилорнитиндезацетилазы из E. coli введены в 10 з.п. ф-лы, 2 ил., 11 табл, 7 пр. R U 2 7 2 1 8 5 2 (56) Список документов, цитированных в отчете о поиске: KR 101053429 B1, 03.08.2011. DE 1642678 A1, 03.06.1971. CN 102191291 A, 21.09.2011. RU 2433180 C2, 10.11.2011. RU 2316588 C1, 10.02.2008. Стр.: 1 C 1 C 1 (45) Опубликовано: 25.05.2020 Бюл. № 15 (73) Патентообладатель(и): СИДЖЕЙ ЧЕИЛДЖЕДАНГ КОРПОРЕЙШН (KR) 2 ...

Microorganisms for producing putrescine or ornithine and method of producing putrescine or ornithine using said microorganisms

Группа изобретений относится к модифицированному микроорганизму рода Corynebacterium, продуцирующему путресцин, и способу получения путресцина с использованием указанного микроорганизма. Предложен модифицированный микроорганизм рода Corynebacterium, в котором активности N-ацетилглутаматсинтазы из E. coli и ацетилорнитиндезацетилазы из E. coli введены в указанный микроорганизм рода Corynebacterium, обладающий активностью ацетилглутаматсинтазы или орнитинацетилтрансферазы (ArgJ). Также предложен способ получения путресцина, включающий культивирование указанного модифицированного микроорганизма рода Corynebacterium в среде и выделение путресцина из культивированного микроорганизма или среды. Группа изобретений позволяет получить путресцин с увеличенным выходом. 2 н. и 15 з.п. ф-лы, 2 ил., 11 табл., 7 пр. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 708 165 C2 (51) МПК C12N 15/77 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/52 (2019.08); C12N 9/1029 (2019.08); C12N 9/1217 (2019.08); C12N 9/80 (2019.08); C12N 9/88 (2019.08); C12N 9/93 (2019.08); C12P 13/001 (2019.08); C12P 13/10 (2019.08); C12Y 203/01001 (2019.08); C12Y 203/01008 (2019.08); C12Y 203/01035 (2019.08); C12Y 207/02001 (2019.08); C12Y 305/01016 (2019.08); C12Y 401/01017 (2019.08); C12Y 602/01001 (2019.08) 2018104512, 19.07.2016 (24) Дата начала отсчета срока действия патента: 19.07.2016 04.12.2019 Приоритет(ы): (30) Конвенционный приоритет: 20.07.2015 KR 10-2015-0102624 (43) Дата публикации заявки: 20.08.2019 Бюл. № 23 (56) Список документов, цитированных в отчете о поиске: US 8497098 D2, 30.07.2013. DOU W. et al. Improvement of L-arginine Production by Overexpression of a Bifunctional Ornithine Acetyltransferase in Corynebacterium Crenatum. Applied Biochemistry and Biotechnology, 2011, vol. 165, pages 845-855. база данных GenBank:EZG54138.1, 09.04.2014. база данных GenBank:BAE77354.1, 20.11.2008. RU 2550269 C2, (см. прод.) (85) Дата начала ...

Microorganisms for obtaining putrescine and method for obtaining putrescine with application thereof

FIELD: chemistry. SUBSTANCE: invention relates to biotechnology, namely to putrescine-producing microorganism and method for obtaining putrescine with thereof application. Putrescine-producing microorganism is modified in such a way that activity of ornithinecarbamoyltransferase and protein, participating in glutamate (NCgl1221) export in comparison with their endogenic activities is deleted in it, and ornithinedecarboxilase (ODC) activity is introduced into microorganism. EFFECT: invention makes it possible to obtain putrescine with high degree of effectiveness. 13 cl, 3 dwg, 8 tbl, 5 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК C12N 1/21 (13) 2 573 923 C2 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2013131033/10, 08.12.2011 (24) Дата начала отсчета срока действия патента: 08.12.2011 Приоритет(ы): (30) Конвенционный приоритет: R U 08.12.2010 KR 10-2010-0124867; 07.12.2011 KR 10-2011-0130595 (72) Автор(ы): ЧОИ Хианг (KR), ЛИ Киоунг Мин (KR), КАНГ Мин Сун (KR), ДЗХОН Сунг Хоо (KR), УМ Хие Вон (KR), ЧОЙ Су Дзин (KR), ЛИ Хан Вон (KR), ШИН Соо Ан (KR) (43) Дата публикации заявки: 20.01.2015 Бюл. № 2 C 2 C 2 (56) Список документов, цитированных в отчете о поиске: SCHNEIDER J et.al. Putrescine production by engineered Corynebacterium glutamicum, Appl Microbiol Biotechnol. 2010 Oct;88(4):859-68. doi: 10.1007/s00253-010-2778-x. Epub 2010 Jul 27. HASHIMOTO K. et.al. The protein encoded by NCgl1221 in Corynebacterium glutamicum functions as a mechanosensitive channel, Biosci Biotechnol Biochem. (см. прод.) (73) Патентообладатель(и): СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 08.07.2013 2 5 7 3 9 2 3 (86) Заявка PCT: R U 2 5 7 3 9 2 3 (45) Опубликовано: 27.01.2016 Бюл. № 3 KR 2011/009478 (08.12.2011) (87) Публикация заявки PCT: WO 2012/077995 (14.06.2012) Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, строение 3, ООО "Юридическая фирма ...

Methods and compositions of otc constructs and vectors

Provided herein are methods and compositions related to nucleic acids encoding ornithine transcarbamylase (OTC), such as nucleic acids comprising an OTC codon- optimized sequence, as well as related vectors, such as AAV vectors. Also, provided are methods for administering AAV vectors that comprise a sequence that encodes an enzyme associated with an urea cycle disorder and an expression control sequence, in combination with synthetic nanocarriers coupled to an immunosuppressant.

Microorganism which produces putrescine or ornithine, and a method of producing putrescine or ornithine using said microorganism

FIELD: biotechnology.SUBSTANCE: disclosed is a modified microorganism of the genus Corynebacterium producing putrescine with reduced activity of the sugar metabolism transcription (SugR) regulator compared to its endogenous activity and high activity of citrate synthase (GltA) compared to its endogenous activity. Disclosed is a method of producing putrescine using said microorganism.EFFECT: group of inventions makes it possible to increase productivity of the specified modified microorganism by putrescine in comparison with non-modified microorganism.15 cl, 7 tbl, 8 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 691 303 C1 (51) МПК C12N 15/77 (2006.01) C12N 9/10 (2006.01) C12P 13/00 (2006.01) C12P 13/10 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/77 (2018.08); C12N 9/1025 (2018.08); C12P 13/001 (2018.08); C12P 13/10 (2018.08); C12Y 203/03001 (2018.08) (21)(22) Заявка: 2017145909, 29.03.2016 29.03.2016 Дата регистрации: 11.06.2019 24.06.2015 KR 10-2015-0090021 (45) Опубликовано: 11.06.2019 Бюл. № 17 (73) Патентообладатель(и): СИДЖЕЙ ЧЕИЛДЖЕДАНГ КОРПОРЕЙШН (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 24.01.2018 (56) Список документов, цитированных в отчете о поиске: JP 2009254323 A, 05.11.2009. KR R U 2 6 9 1 3 0 3 C 1 KR 2016/003198 (29.03.2016) (87) Публикация заявки PCT: WO 2016/208854 (29.12.2016) Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" (54) Микроорганизм, продуцирующий путресцин или орнитин, и способ получения путресцина или орнитина с использованием этого микроорганизма (57) Реферат: Группа изобретений относится к области с ее эндогенной активностью. Предложен способ биотехнологии. Предложен модифицированный получения путресцина с использованием микроорганизм рода Corynebacterium, указанного микроорганизма. Группа изобретений продуцирующий путресцин, с пониженной позволяет увеличить продуктивность указанного активностью регулятора транскрипции ...

Putrescine or ornithine producing microorganism and method for producing putrescine or ornithine using the same

本発明はプトレシンまたはオルニチンを生産するための組み換え微生物及びそれを用いてプトレシンまたはオルニチンを生産する方法に関する。 【選択図】図２

Microorganism producing putrescine or ornithine and method for producing putrescine or ornithine using this microorganism

FIELD: microbiology.SUBSTANCE: group of inventions relates to a modified microorganism for producing ornithine and to a method for producing ornithine using this microorganism. A modified microorganismCorynebacterium glutamicumis proposed, producing ornithine with reduced activity of a regulator of sugar metabolism transcription (SugR) in comparison with endogen activity ofCorynebacterium glutamicumand increased activity of citrate synthase (GltA) in comparison with endogen activity ofCorynebacterium glutamicum, where thisCorynebacterium glutamicumhas a capability of producing ornithine. A method for producing ornithine using the specified microorganism is also proposed.EFFECT: group of inventions provides producing ornithine with high yield.8 cl, 7 tbl, 8 ex РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 759 956 C1 (51) МПК C12N 15/77 (2006.01) C12N 9/10 (2006.01) C12N 9/88 (2006.01) C12P 13/10 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ ИЗОБРЕТЕНИЯ К ПАТЕНТУ (52) СПК C12N 15/77 (2021.08); C12N 9/1018 (2021.08); C12N 9/1025 (2021.08); C12N 9/88 (2021.08); C12P 13/10 (2021.08); C12Y 201/03003 (2021.08); C12Y 203/03001 (2021.08); C12Y 401/01017 (2021.08) (21)(22) Заявка: 2019103640, 29.03.2016 29.03.2016 Дата регистрации: 19.11.2021 24.06.2015 KR 10-2015-0090021 Номер и дата приоритета первоначальной заявки, из которой данная заявка выделена: 2017145909 24.06.2015 C 1 2 7 5 9 9 5 6 Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" (54) Микроорганизм, продуцирующий путресцин или орнитин, и способ получения путресцина или орнитина с использованием этого микроорганизма (57) Реферат: Группа изобретений относится к glutamicum и повышенной активностью модифицированному микроорганизму для цитратсинтазы (GltA) по сравнению с эндогенной получения орнитина и к способу получения активностью Corynebacterium glutamicum, где этот орнитина с использованием этого Corynebacterium glutamicum обладает микроорганизма. Предложен модифицированный ...

MICROORGANISMS WITH THE IMPROVED ORNITINE-PRODUCING ABILITY, AND METHOD OF OBTAINING ORNITINE USING THEM

1. Микроорганизм, имеющий улучшенную орнитин-продуцирующую способность, где активности орнитинкарбамоилтрансферазы и белка, участвующего в экспорте глутамата (NCgl1221), модифицированы таким образом, что они являются аттенуированными в сравнении с их эндогенными активностями.2. Микроорганизм по п. 1, где орнитинкарбамоилтрансфераза имеет аминокислотную последовательность SEQ ID NO:18 или аминокислотную последовательность, имеющую 70% или более высокую гомологию с этой последовательностью.3. Микроорганизм по п. 1, где белок, участвующий в экспорте глутамата, имеет аминокислотную последовательность SEQ ID NO:20 или аминокислотную последовательность, имеющую 70% или более высокую гомологию с этой последовательностью.4. Микроорганизм по п. 1, где активность орнитинкарбамоилтрансферазы и белка, участвующего в экспорте глутамата, является аттенуированной способом, выбранным из группы, состоящей из (1) частичной или полной делеции гена, кодирующего этот белок, (2) модификации регуляторной последовательности экспрессии для супрессии экспрессии гена, (3) модификации последовательности гена на хромосоме для уменьшения активности этого белка и (4) их комбинации.5. Микроорганизм по п. 1, где активности ацетил-гамма-глутамилфосфатредуктазы (ArgC), ацетилглутаматсинтазы или орнитинацетилтрансферазы (ArgJ), ацетилглутаматкиназы (ArgB) и ацетилорнитинаминотрансферазы (ArgD) являются дополнительно усиленными в сравнении с их эндогенными активностями.6. Микроорганизм по п. 5, где каждый из ArgC, ArgJ, ArgB и ArgD имеет аминокислотную последовательность SEQ ID NO:23, 25, 27 и 29 или аминокислотную последовательность, имеющую 70% или более высокую гомологию с этой послед� РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (51) МПК C12N 1/21 (13) 2013 131 039 A (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2013131039/10, 08.12.2011 (71) Заявитель(и): СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН (KR) Приоритет(ы): (30) Конвенционный приоритет: 08.12.2010 KR ...

Microorganisms for producing putrescine or ornithine and process for producing putrescine or ornithine using them

본 발명은 퓨트레신 또는 오르니틴을 생산하기 위한 재조합 미생물 및 이를 이용하여 퓨트레신 또는 오르니틴을 생산하는 방법에 관한 것이다. The present invention relates to a recombinant microorganism for producing putrescine or ornithine, and a method for producing putrescine or ornithine using the recombinant microorganism.

Patent RU2018104512A3

РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2018 104 512 A (51) МПК C12N 15/77 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2018104512, 19.07.2016 (71) Заявитель(и): СИДЖЕЙ ЧЕИЛДЖЕДАНГ КОРПОРЕЙШН (KR) Приоритет(ы): (30) Конвенционный приоритет: 20.07.2015 KR 10-2015-0102624 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.02.2018 (86) Заявка PCT: (87) Публикация заявки PCT: WO 2017/014532 (26.01.2017) R U (54) Микроорганизмы для получения путресцина или орнитина и способ получения путресцина или орнитина с использованием указанных микроорганизмов (57) Формула изобретения 1. Модифицированный микроорганизм рода Corynebacterium, продуцирующий путресцин или орнитин, в который введены активности N-ацетилглутаматсинтазы из Е. coli и ацетилорнитиндезацетилазы из Е. coli. 2. Модифицированный микроорганизм по п. 1, где N-ацетилглутаматсинтаза из Е. coli состоит из аминокислотной последовательности SEQ ID NO: 1. 3. Модифицированный микроорганизм по п. 1, где ацетилорнитиндезацетилаза из Е. coli состоит из аминокислотной последовательности SEQ ID NO: 3. 4. Модифицированный микроорганизм по п. 1, где микроорганизм рода Corynebacterium представляет собой Corynebacterium glutamicum. 5. Модифицированный микроорганизм по п. 1, где активность фосфотрансацетилазного и ацетаткиназного оперона (оперона pta-ackA) дополнительно усилена по сравнению с его эндогенной активностью. 6. Модифицированный микроорганизм по п. 5, где фосфотрансацетилазный и ацетаткиназный оперон состоит из аминокислотной последовательности SEQ ID NO: 5 или SEQ ID NO: 7. 7. Модифицированный микроорганизм по п. 1, в который дополнительно введена активность ацетил-КоА-синтетазы (acs) из Е. coli. 8. Модифицированный микроорганизм по п. 7, где ацетил-КоА-синтетаза состоит из аминокислотной последовательности SEQ ID NO: 9. Стр.: 1 A 2 0 1 8 1 0 4 5 1 2 A Адрес для переписки: 191036, Санкт-Петербург, а/я 24, "НЕВИНПАТ" 2 0 1 8 1 0 4 5 1 2 KR 2016/ ...

Микроорганизмы, имеющие улучшенную орнитин-продуцирующую способность, и способ получения орнитина с их использованием

Изобретение относится к биотехнологии и представляет собой микроорганизм, имеющий орнитин-продуцирующую способность, где активности орнитинкарбамоилтрансферазы и белка, участвующего в экспорте глутамата (NCgl1221), модифицированы таким образом, что они являются делетированными в сравнении с их эндогенными активностями. Изобретение относится также к способу получения орнитина с использованием этого микроорганизма. Изобретение позволяет получать L-орнитин с высокой степенью эффективности. 2 н. и 9 з.п. ф-лы, 3 ил., 6 табл., 4 пр. РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2 567 669 C2 (51) МПК C12N 1/21 (2006.01) C12P 13/04 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ОПИСАНИЕ (21)(22) Заявка: ИЗОБРЕТЕНИЯ К ПАТЕНТУ 2013131039/10, 08.12.2011 (24) Дата начала отсчета срока действия патента: 08.12.2011 Приоритет(ы): (30) Конвенционный приоритет: R U 08.12.2010 KR 10-2010-0124866; 07.12.2011 KR 10-2011-0130594 (72) Автор(ы): ЧОИ Хианг (KR), ЛИ Киоунг Мин (KR), КАНГ Мин Сун (KR), ДЗХОН Сунг Хоо (KR), УМ Хие Вон (KR), ЧОЙ Су Дзин (KR), ЛИ Хан Вон (KR), ШИН Соо Ан (KR) (43) Дата публикации заявки: 20.01.2015 Бюл. № 2 C 2 C 2 (56) Список документов, цитированных в отчете о поиске: SCHNEIDER J et.al. Production of the amino acids l-glutamate, l-lysine, l-ornithine and l-arginine from arabinose by recombinant Corynebacterium glutamicum, J Biotechnol. 2011 Jul 10;154(2-3):191-8. doi: 10.1016/ j.jbiotec.2010.07.009. Epub 2010 Jul 16.RU 2316588 C1, 10.02.2008.HASHIMOTO K. et.al. The protein encoded by NCgl1221 in Corynebacterium (см. прод.) (73) Патентообладатель(и): СиДжей ЧЕИЛДЗЕДАНГ КОРПОРЕЙШН (KR) (85) Дата начала рассмотрения заявки PCT на национальной фазе: 08.07.2013 2 5 6 7 6 6 9 (86) Заявка PCT: R U 2 5 6 7 6 6 9 (45) Опубликовано: 10.11.2015 Бюл. № 31 KR 2011/009477 (08.12.2011) (87) Публикация заявки PCT: WO 2012/077994 (14.06.2012) Адрес для переписки: 129090, Москва, ул. Б. Спасская, 25, строение 3, ООО "Юридическая фирма Городисский и Партнеры" (54 ...

Putrescine-producing microorganisms and a method for producing putrescine using these microorganisms

РОССИЙСКАЯ ФЕДЕРАЦИЯ (19) RU (11) (13) 2015 142 261 A (51) МПК C12N 1/21 (2006.01) ФЕДЕРАЛЬНАЯ СЛУЖБА ПО ИНТЕЛЛЕКТУАЛЬНОЙ СОБСТВЕННОСТИ (12) ЗАЯВКА НА ИЗОБРЕТЕНИЕ (21)(22) Заявка: 2015142261, 25.02.2014 (71) Заявитель(и): СиДжей ЧеилДжеданг Корпорейшн (KR) Приоритет(ы): (30) Конвенционный приоритет: 20.03.2013 KR 10-2013-0030020; 14.02.2014 KR 10-2014-0017243 (85) Дата начала рассмотрения заявки PCT на национальной фазе: 20.10.2015 KR 2014/001509 (25.02.2014) (87) Публикация заявки PCT: WO 2014/148743 (25.09.2014) R U (54) Микроорганизмы, продуцирующие путресцин, и способ получения путресцина с использованием этих микроорганизмов (57) Формула изобретения 1. Микроорганизм, продуцирующий путресцин, где указанный микроорганизм модифицирован для усиления активности белка, имеющего аминокислотную последовательность, представленную в SEQ ID NO: 21 или 23. 2. Микроорганизм, продуцирующий путресцин, по п. 1, дополнительно модифицированный для ослабления активности орнитинкарбамоилтрансферазы (ArgF) и белка (NCgl1221), вовлеченного в экспорт глутамата, по сравнению с эндогенной активностью и для усиления активности орнитиндекарбоксилазы (ODC). 3. Микроорганизм, продуцирующий путресцин, по п. 2, где орнитинкарбамоилтрансфераза (ArgF) имеет аминокислотную последовательность, представленную в SEQ ID NO: 29, белок (NCgl1221) вовлеченный в экспорт глутамата, имеет аминокислотную последовательность, представленную в SEQ ID NO: 30, и орнитиндекарбоксилаза (ODC) имеет аминокислотную последовательность, представленную в SEQ ID NO: 33. 4. Микроорганизм, продуцирующий путресцин, по п. 1, дополнительно модифицированный для усиления активности ацетил-гамма-глутамилфосфатредуктазы (ArgC), ацетилглутаматсинтазы или орнитинацетилтрансферазы (ArgJ), ацетилглутаматкиназы (ArgB) и ацетилорнитинаминотрансферазы (ArgD) по сравнению с эндогенной активностью. 5. Микроорганизм, продуцирующий путресцин, по п. 4, где ацетил-гаммаСтр.: 1 A 2 0 1 5 1 4 2 2 6 1 A Адрес для переписки: 191036, Санкт- ...

mRNA for use in treatment of human genetic diseases

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification conferring stability to the RNA, as well as related methods, are disclosed.

Novel polypeptide and a method for producing a desired product derived from ornithine using the same

본 출원은 오르니틴계 산물 배출능을 가지는 신규한 폴리펩타이드 및 이를 이용하여 오르니틴계 산물을 생산하는 방법에 관한 것이다. The present invention relates to a novel polypeptide having ornithine-based product excretion ability and a method for producing the ornithine product using the same.

Recombinant microorganism and method for producing orotic acid

本发明提供了多种生产乳清酸的重组微生物以及利用重组微生物生产乳清酸的方法，这些重组微生物菌种含有一个或几个下列特征或全部特征，这些特征是：（1）乳清酸代谢途径中的磷酸核糖转移酶或乳清酸核苷单磷酸脱羧酶的合成减弱了或完全丧失了；（2）通过超表达参与乳清酸合成途径中的酶的五个基因来增加相应的酶量；（3）通过超表达从胞内转运乳清酸的转运蛋白编码基因来增加相应的转运蛋白。

MULTIMER CODING NUCLEIC ACID AND ITS USES

Improved composition and methods for treatment of ornithine transcarbamylase deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase protein at a low dose and at an administration interval such that at least one symptom or feature of the OTC deficiency is reduced.

How to choose nutritional requirements

본 개시내용은 영양요구성 세포의 집단 및 분화된 세포 집단을 생성하고, 영양요구성 분할을 사용하여 형질감염된 세포의 집단을 선택하기 위한 방법 및 조성물을 제공한다.

Auxotrophic selection methods

The present disclosure provides methods and compositions for generating populations of auxotrophic cells and populations of differentiated cells and selecting populations of transfected cells using split auxotrophy.

Cleavable lipids

Disclosed herein are novel compounds, pharmaceutical compositions comprising such compounds and related methods of their use. The compounds described herein are useful, e.g. as liposomal delivery vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and subsequent transfection of said target cells, and in certain embodiments are characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids.

Method for the fermentative preparation of L-amino acids and nucleotide sequences coding for the accDA gene

Arginine supplementation to improve efficiency in gas fermenting acetogens

The invention provides methods for improving efficiency of fermentation by arginine supplementation, and genetically modified bacterium for use therefor. More particularly the invention provides methods for (i) increasing the production ATP intensive products with arginine supplementation, (ii) increasing utilization of arginine by a C1-fixing bacterium; and (iii) providing C1-fixing bacterium with optimized arginine de-aminase pathways.

Cleavable Lipids

Disclosed herein are novel compounds, pharmaceutical compositions comprising such compounds and related methods of their use. The compounds described herein are useful, e.g., as liposomal delivery vehicles to facilitate the delivery of encapsulated polynucleotides to target cells and subsequent iransfection of said target cells, and in certain embodiments are characterized as having one or more properties that afford such compounds advantages relative to other similarly classified lipids.

MESSENGER RNA PURIFICATION PROCESSES

mRNA FOR USE IN TREATMENT OF HUMAN GENETIC DISEASES

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification conferring stability to the RNA, as well as related methods, are disclosed.

Method for synthesizing orotic acid by biological enzyme method

本发明公开一种生物酶法合成乳清酸的方法，以氨甲酰磷酸合成酶、天冬氨酸转氨甲酰酶、二氢乳清酸合成酶、二氢乳清酸脱氢酶组成体外多酶催化体系，催化底物转化为乳清酸，所述底物包括谷氨酰胺和/或氨、CO 2 和/或碳酸氢盐、天冬氨酸、ATP和/或ADP，和其它少量无机盐。本发明构建的方法实现了在体外利用纯酶反应以碳酸氢盐和天冬氨酸等为原料合成乳清酸，通过对反应温度、pH、酶量和底物浓度等进行优化和控制，提高了体外合成乳清酸的能力，可以避免微生物发酵法带来的副产物多，后续分离工艺等不利影响。体外纯酶反应可以缩短生产周期，本发明仅需要2‑5h即可获得产物，而微生物发酵法则需要72h，本发明有效缩短反应时间。

Auxotrophy selection method

本公开内容提供了用于产生营养缺陷型细胞群体和分化细胞群体以及使用拆分营养缺陷型选择转染细胞群体的方法和组合物。

Putrescine or ornithine producing microorganism and method for producing putrescine or ornithine using same

The present invention relates to a recombinant microorganism for producing putrescine or ornithine and a method for producing putrescine or ornithine using the same. Specifically, the present invention relates to: a

Process for the preparation of l-amino acids by fermentation and nucleotide sequences coding for the accda gene

The invention relates to nucleotide sequences coding for the accDA gene and to a process for the preparation of L-amino acids, especially L-lysine, by fermentation using corynebacteria in which the accDA gene is amplified.

Method for the fermentative preparation of l-amino acids and nucleotide sequences coding for the accda gene

Cloned Corynebacterium glutamicum DNA (I), replicable in coryneform microorganisms coding for an accDA gene having a fully defined 2123 bp sequence (given in the specification(), is new. Independent claims are also included for the following: (1) a polypeptide derived from (I) having a fully defined 491 aa sequence (given in the specification); (2) coryneform microorganisms transformed with one or more copies of (I); (3) the shuttle vector pZ1accDA contained in Corynebacterium glutamicum DSM 12785; and (4) a process for producing L-amino acids, comprising culturing a coryneform bacterium that overexpresses the accDA gene.

Fusion proteins

Methods and compositions of otc constructs and vectors

Provided herein are methods and compositions related to nucleic acids encoding ornithine transcarbamylase (OTC), such as nucleic acids comprising an OTC codon- optimized sequence, as well as related vectors, such as AAV vectors. Also, provided are methods for administering AAV vectors that comprise a sequence that encodes an enzyme associated with an urea cycle disorder and an expression control sequence, in combination with synthetic nanocarriers coupled to an immunosuppressant.

Microorganisms for producing putrescine or ornithine and process for producing putrescine or ornithine using them

The present invention relates to a recombinant microorganism for producing putrescine or ornithine, and a method for producing putrescine or ornithine using the same. Specifically, the present invention relates to a microorganism of the genus Corynebacterium capable of producing putrescine or ornithine, in which an activity of the transcriptional regulator of sugar metabolism (SugR) is weakened, an activity of the citrate synthase (GltA) is enhanced, or both are applied; and a method for producing putrescine or ornithine using the same.

Methods for purification of messenger rna

The present invention relates, in part, to methods for large-scale purification of mRNA. The method includes, at least, a step of centrifuging an mRNA suspension in a centrifuge comprising a porous substrate at a speed sufficient to remove process contaminants and to precipitate purified mRNA composition onto the porous substrate.

Transformed methanotrophs for producing 3-Hydroxypropionic acid and uses thereof

본 발명은 말로닐-CoA 환원 효소(malonyl-CoA reductase); 아세틸-CoA 카르복실화 효소(acetyl-CoA carboxylase) 및/또는 메틸말로닐-CoA 카르복실전달효소(Methylmalonyl-CoA carboxyltransferase) 활성이 강화된, 3-하이드록시 프로피온산(3-Hydroxypropionic acid) 생산용 형질전환 메탄자화균; 상기 형질전환 메탄자화균을 사용한 3-하이드록시프로피온산의 제조방법; 및 상기 형질전환 메탄자화균을 포함하는 3-하이드록시프로피온산 생산용 조성물에 관한 것이다. 본 발명의 형질전환 메탄자화균은 C1 탄소원으로부터 3-하이드록시프로피온산을 높은 수율로 생산할 수 있는바, 친환경적이고 경제적으로 3-하이드록시프로피온산의 대량 생산에 유용하게 사용될 수 있다.

Adeno-associated virus (AAV) serotype 8 sequences, vectors containing same, and uses therefor

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles.

Polymer code nucleic acid and application thereof

本发明尤其提供了多聚体编码核酸，其显示出用于体内和体外使用的优异稳定性。在一些实施例中，多聚体编码核酸(MCNA)包含经由3'末端连接的两种或更多种编码多核苷酸，使得多聚体编码核酸化合物包含两个或更多个5'末端。

Improved composition and methods for treatment of ornithine transcarbamylase deficiency

The present invention provides, among other things, methods of treating ornithine transcarbamylase deficiency, including administering to a subject in need of treatment a composition comprising an mRNA encoding an ornithine transcarbamylase protein at a low dose and at an administration interval such that at least one symptom or feature of the OTC deficiency is reduced.

mRNA for use in treatment of human genetic diseases

Compositions for modulating the expression of a protein in a target cell comprising at least one RNA molecule which comprises at least one modification 5 conferring stability to the RNA, as well as related methods, are disclosed.

Tricine and citric acid lipids

Disclosed are cationic lipids which are compounds of Formula (A). Cationic lipids provided herein can be useful for delivery and expression of mRNA and encoded protein, e.g., as a component of liposomal delivery vehicle, and accordingly can be useful for treating various diseases, disorders and conditions, such as those associated with deficiency of one or more proteins.

Microorganisms having putrescine productivity and process for producing putrescine using the same

本發明係關於一種能夠產生腐胺之重組微生物，其中該微生物經修飾以具有增強的NCgl2522活性，藉此以高產率產生腐胺；以及一種使用該微生物產生腐胺之方法。

Stable gene transfer to proliferating cells

Provided herein are methods for facilitating or inducing stable transgene integration and expression in a proliferating cell, comprising administering to the cell (i) a recombinant AAV (rAAV) vector comprising the transgene flanked by transposon-derived inverted terminal repeat sequences, which sequences are in turn flanked by AAV-derived inverted terminal repeat regions, and (ii) a source of a transposase that recognises said transposon-derived inverted terminal repeat sequences and directs the genomic integration of the transgene into the genome of the proliferating cell. Also provided are methods and transgene delivery systems for the treatment or prevention of diseases affecting, associated with or characterised by proliferating cells, including paediatric liver diseases, bone marrow diseases and cancer.

Compositions and methods for treating ornithine transcarbamylase deficiency

The present disclosure provides a modified human OTC protein having improved properties for the treatment of OTC deficiency in a patient. Preferably, the protein of the disclosure is produced from a codon optimized mRNA suitable for administration to a patient suffering from OTC deficiency wherein upon administration of the mRNA to the patient, the protein of the disclosure is expressed in the patient in therapeutically effective amounts to treat OTC deficiency. The present disclosure also provides codon optimized mRNA sequences encoding wild type human OTC comprising a 5' UTR derived from a gene expressed by Arabidopsis thaliana for use in treating OTC deficiency in a patient.

Adeno-associated virus (aav) serotype 8 sequences, vectors containing same, and uses therefor

Sequences of a serotype 8 adeno-associated virus and vectors and host cells containing these sequences are provided. Also described are methods of using such host cells and vectors in production of rAAV particles.