Light-responsive polypeptides and methods of use thereof

Patent metadata

Field	Value
Publication number	US-11801301-B2
Application number	US-201615756936-A
Country	US
Kind code	B2
Filing date	Sep 14, 2016
Priority date	Sep 15, 2015
Publication date	Oct 31, 2023
Grant date	Oct 31, 2023

Abstract

The present disclosure provides variant light-responsive polypeptides, and nucleic acids comprising nucleotide sequences encoding the light-responsive polypeptides. The present disclosure provides methods, devices, and systems for controlling the activity of a cell expressing a variant light-responsive polypeptide of the present disclosure.

First claim

What is claimed is:

1. A variant light-activated polypeptide that comprises an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:3, wherein the amino acid sequence comprises serine at position 123 relative to the amino acid sequence set forth in SEQ ID NO:3, and wherein the variant light-activated polypeptide exhibits at least 5-fold increased channel closure kinetics compared to a light-activated polypeptide of SEQ ID NO:1.

2. The variant light-activated polypeptide of claim 1 , comprising:
a) a heterologous membrane trafficking signal;
b) an endoplasmic reticulum (ER) export signal; or
c) both a heterologous membrane trafficking signal and an ER export signal.

3. The variant light-activated polypeptide of claim 1 , wherein the variant light-activated polypeptide comprises an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in one of SEQ ID NO:4, SEQ ID NO:5, and SEQ ID NO:6.

4. The variant light-activated polypeptide of claim 1 , wherein variant light-activated polypeptide evokes action potentials at frequencies of from 5 Hz to 20 Hz when present in a eukaryotic cell and when activated by light of a wavelength of from 600 nm to 700 nm.

5. A nucleic acid comprising a nucleotide sequence encoding the variant light-activated polypeptide of claim 1 .

6. A recombinant expression vector comprising the nucleic acid of claim 5 .

7. A mammalian cell comprising the variant light-activated polypeptide of claim 1 in the cell membrane, wherein the variant light-activated polypeptide is responsive to light, wherein the variant light-activated polypeptide is capable of mediating a depolarizing current in the cell when the cell is illuminated with light of a wavelength of from about 600 nm to about 700 nm.

8. The variant light-activated polypeptide of claim 1 , wherein the variant light-activated polypeptide exhibits at least 10-fold increased channel closure kinetics compared to the light-activated polypeptide of SEQ ID NO:1.

Assignees

• Univ Leland Stanford Junior

Inventors

• Deisseroth Karl A
• Berndt Andre
• Lee Soo Yeun
• Ramakrishnan Charu

Classifications

• A61K41/00 Primary

  Medicinal preparations obtained by treating materials with wave energy or particle radiation {; Therapies using these preparations} · CPC title

• A61N5/06

  using light · CPC title

• A61N5/062

  Photodynamic therapy, i.e. excitation of an agent · CPC title

• C07K14/00

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof · CPC title

• C07K14/405

  from algae · CPC title