Opsin polypeptides and methods of use thereof
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsKarl Deisseroth, Feng Zhang, Viviana Gradinaru
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Abstract
The present disclosure provides opsins, including variant opsins with increased activity and/or increased trafficking to the plasma membrane. The opsins are useful in therapeutic and screening applications, which are also provided.
Claims14
An isolated polynucleotide comprising a nucleotide sequence encoding a fusion polypeptide comprising, in order from amino terminus to carboxyl terminus:
- a) an opsin polypeptide comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:33;
- b) a membrane trafficking signal comprising the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:
- 10);
- and
- c) an endoplasmic reticulum (ER) export signal comprising the amino acid sequence FXYENE (SEQ ID NO:
- 4), where X is any amino acid.
A mammalian cell comprising a light-activated fusion polypeptide expressed on the cell membrane, wherein the light-activated fusion polypeptide is capable of mediating a hyperpolarizing current in the cell when the cell is illuminated with light, and wherein the light-activated fusion polypeptide comprises, in order from amino terminus to carboxyl terminus:
- a) an opsin polypeptide comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:33;
- b) a membrane trafficking signal comprising the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:
- 10);
- and
- c) an endoplasmic reticulum (ER) export signal comprising the amino acid sequence FXYENE (SEQ ID NO:
- 4), where X is any amino acid.
A method of modulating the voltage potential of a cell in response to a light stimulus, the method comprising exposing a cell to a light stimulus, wherein the cell that is genetically modified with a recombinant expression vector comprising a nucleotide sequence encoding a fusion polypeptide, wherein the fusion polypeptide comprises, in order from amino terminus to carboxyl terminus:
- i) an opsin polypeptide comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:33;
- ii) a membrane trafficking signal comprising the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:
- 10);
- and
- ii) an endoplasmic reticulum (ER) export signal comprising the amino acid sequence FXYENE (SEQ ID NO:
- 4), where X is any amino acid,
- wherein the encoded fusion polypeptide is expressed in the cell, and wherein, in response to exposure to a light stimulus, the voltage potential of the cell is modulated.
Dependencies are inferred from the claim text.