14-03-2013 дата публикации
Номер: US20130066402A1
The invention provides engineered red-shifted channelrhodopsin variants. In some embodiments, the channelrhodopsin variants are characterized by improved membrane trafficking, expression, and/or unique spectral and kinetic properties. 1Volvox carteriVolvox carteri. A polypeptide comprising a channelrhodopsin-1 (ChR1) domain , a channelrhodopsin-1 (VChR1) domain and a channelrhodopsin-2 (VChR2) domain.2. The polypeptide of claim 1 , wherein the polypeptide has the structure:{'sup': 1', '2', '3', '4, 'X-X-X-X'}{'sup': '1', 'wherein Xis a ChR1 domain,'}{'sup': '2', 'Xis a first VChR1 domain,'}{'sup': '3', 'Xis a VChR2 domain and'}{'sup': '4', 'Xis a second VChR1 domain.'}3. The polypeptide of claim 1 , having a sequence according to SEQ ID NO: 1.4. The polypeptide of claim 2 , wherein one or more of the domains X claim 2 , X claim 2 , X claim 2 , and Xcomprises 1 claim 2 , 2 claim 2 , 3 claim 2 , 4 claim 2 , or 5 amino acid substitutions relative to a corresponding wild-type domain.5. The polypeptide of claim 4 , wherein the amino acid substitution is at one or more positions selected from 163 claim 4 , 171 claim 4 , 174 and 266.6. The polypeptide of claim 4 , comprising one or more amino acid substitutions selected from Glu163Thr claim 4 , Leu171Ile claim 4 , Leu171Val claim 4 , His174Arg claim 4 , and Phe266Tyr.7. The polypeptide of further comprising a fluorescent polypeptide.8. A polynucleotide encoding the polypeptide of .9. The polynucleotide of claim 8 , having a sequence according to SEQ ID NO: 2.10. A method of depolarizing a cell comprising:{'claim-ref': {'@idref': 'CLM-00001', 'claim 1'}, '(a) expressing the polypeptide of within the cell; and'}(b) exposing the cell to light.11. (canceled)12. (canceled)13. A method of restoring sensitivity to light in an ocular cell comprising expressing the polypeptide of in the ocular cell.14. The polypeptide of claim 1 , comprising an amino acid sequence that is at least 85% identical to SEQ ID NO:1.15. The polypeptide of ...
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