Light-activated chimeric opsins and methods of using the same
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsKarl Deisseroth, Ofer Yizhar, Lief Fenno, Peter Hegemann, Matthias Prigge
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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.
Claims21
A method for identifying a chemical compound that selectively inhibits the depolarization of excitatory or inhibitory neurons residing in the same microcircuit, the method comprising:
- (a) selectively depolarizing an excitatory neuron comprising a first light-activated protein with light having a first wavelength or selectively depolarizing an inhibitory neuron comprising a second light-activated protein with light having a second wavelength;
- wherein the first or the second light activated protein is a chimeric protein 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;
- (b) measuring an excitatory post synaptic potential (EPSP) in response to selectively depolarizing the excitatory neuron comprising the first light-activated protein or measuring an inhibitory post synaptic current (IPSC) in response to selectively depolarizing an inhibitory neuron comprising the second light-activated protein;
- (c) contacting the excitatory neuron or the inhibitory neuron with a chemical compound;
- and
- (d) measuring the excitatory post synaptic potential (EPSP) or measuring the inhibitory post synaptic current (IPSC) to determine if contacting either the excitatory neuron or the inhibitory neuron with the chemical compound selectively inhibits the depolarization of either neuron.
A light-responsive chimeric polypeptide comprising, in order from N-terminus to C-terminus:
- a) an amino acid sequence having at least 90% amino acid sequence identity to amino acids 1-163 of the ChR1 amino acid sequence set forth in SEQ ID NO:16;
- and
- b) an amino acid sequence having at least 90% amino acid sequence identity to amino acids 120-300 of SEQ ID NO:17.
A polynucleotide comprising a nucleotide sequence encoding a light-responsive chimeric polypeptide comprising, in order from N-terminus to C-terminus:
- a) an amino acid sequence having at least 90% amino acid sequence identity to amino acids 1-163 of the ChR1 amino acid sequence set forth in SEQ ID NO:
- 16;
- and
- b) an amino acid sequence having at least 90% amino acid sequence identity to amino acids 120-300 of SEQ ID NO:
- 17.
Dependencies are inferred from the claim text.