Light-activated chimeric opsins and methods of using the same

Patent metadata

Field	Value
Publication number	US-9340589-B2
Application number	US-201514862926-A
Country	US
Kind code	B2
Filing date	Sep 23, 2015
Priority date	Nov 5, 2010
Publication date	May 17, 2016
Grant date	May 17, 2016

Abstract

Provided herein are compositions comprising light-activated chimeric proteins expressed on plasma membranes and methods of using the same to selectively depolarize excitatory or inhibitory neurons.

First claim

What is claimed is:

1. A mammalian cell comprising a polynucleotide comprising a nucleotide sequence encoding a light-responsive chimeric polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:1, wherein the light-responsive chimeric polypeptide is present in the cell membrane.

2. The mammalian cell of claim 1 , wherein the light-responsive chimeric polypeptide comprises a C-terminal trafficking signal.

3. The mammalian cell of claim 2 , wherein the trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:15).

4. The mammalian cell of claim 1 , wherein the nucleotide sequence is operably linked to a promoter.

5. The mammalian cell of claim 1 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 122 relative to the amino acid sequence set forth in SEQ ID NO:1.

6. The mammalian cell of claim 1 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 162 relative to the amino acid sequence set forth in SEQ ID NO:1.

7. The mammalian cell of claim 1 , wherein the light-responsive chimeric polypeptide comprises a Glu to Thr amino acid substitution at position 122 and a Glu to Thr amino acid substitution at position 162 relative to the amino acid sequence set forth in SEQ ID NO:1.

8. The mammalian cell of claim 1 , wherein the cell is a neuronal cell, a muscle cell, or a stem cell.

9. The mammalian cell of claim 1 , wherein the cell is an excitatory neuronal cell or an inhibitory neuronal cell.

10. The mammalian cell of claim 1 , wherein the cell further comprises a second light-activated polypeptide present in the cell membrane.

11. The mammalian cell of claim 10 , wherein the second light-activated polypeptide comprises an amino acid sequence having at least 85% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NOs: 11, 12, 13, or 14.

12. A method of depolarizing an excitatory or an inhibitory neuron, the method comprising exposing the neuron to light of a wavelength between about 540 nm to about 560 nm, wherein the neuron expresses in its cell membrane a light-responsive chimeric polypeptide comprising an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:1.

13. The method of claim 12 , wherein the neuron is an excitatory neuron present in a microcircuit, and where the method further comprises expressing a second light-activated polypeptide in an inhibitory neuron, wherein the second light-activated polypeptide comprises an amino acid sequence having at least 85% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NOs: 11, 12, 13, or 14, and wherein the inhibitory neuron is exposed to light of a second wavelength.

14. The method of claim 12 , wherein the excitatory neuron is present in the prefrontal cortex.

15. The method of claim 12 , wherein the excitatory neuron comprises a pyramidal neuron.

16. The method of claim 12 , wherein the inhibitory neuron is present in the prefrontal cortex.

17. The method of claim 12 , wherein the inhibitory neuron comprises a parvalbumin neuron.

18. The method of claim 13 , wherein the excitatory neuron and the inhibitory neuron are present in the prefrontal cortex.

19. The method of claim 13 , wherein the excitatory neuron and the inhibitory neuron are present in the prefrontal cortex.

Assignees

• Univ Leland Stanford Junior

Inventors

• Deisseroth Karl
• Yizhar Ofer
• Fenno Lief
• Hegemann Peter
• Prigge Matthias

Classifications

• A61P25/18

  Antipsychotics, i.e. neuroleptics; Drugs for mania or schizophrenia · CPC title

• A61P25/28

  for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia · CPC title

• A61P25/00

  Drugs for disorders of the nervous system · CPC title

• C07K2319/00

  Fusion polypeptide · CPC title

• A61N2005/0663

  Coloured light · CPC title