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.
Claims51
An animal cell comprising a light-activated protein expressed on the cell membrane, wherein the protein
- is a chimeric protein derived from VChR1 from Volvox carteri and ChR1 from Chlamydomonas reinhardti, wherein the protein comprises the amino acid sequence of VChR1 having at least the first and second transmembrane helices replaced by the first and second transmembrane helices of ChR1;
- is responsive to light;
- and
- is capable of mediating a depolarizing current in the cell when the cell is illuminated. with light.
An isolated polynucleotide comprising a nucleotide sequence encoding a light activated protein expressed on the cell membrane, wherein the protein is a chimeric protein derived from VChR1 from Volvox carteri and ChR1 from Chlamydomonas reinhardti, wherein the protein comprises the amino acid sequence of VChR1 having at least the first and second transmembrane helices replaced by the first and second transmembrane helices of ChR1;
- is responsive to light;
- and is capable of mediating a depolarizing current in the cell when the cell is illuminated with light.
A method of selectively depolarizing excitatory or inhibitory neurons residing in the same microcircuit, the method comprising:
- selectively depolarizing an excitatory neuron comprising a first light-activated protein, wherein the first light activated protein is depolarized when exposed to light having a first wavelength;
- or selectively depolarizing an inhibitory neuron comprising a second light-activated protein, wherein the second light activated protein is depolarized when exposed to light having a second wavelength:, wherein the first or the second light activated protein is a chimeric protein derived from VChR1 from Volvox carteri and ChR1 from Chlamydomonas reinhardti, wherein the protein comprises the amino acid sequence of VChR1 having at least the first and second transmembrane helices replaced by the first and second transmembrane helices of ChR1.
A method of selectively depolarizing excitatory or inhibitory neurons residing in the same microcircuit, the method comprising:
- expressing a first light-activated protein in an excitatory neuron;
- and expressing a second light activated protein in an inhibitory neuron, wherein the first light activated protein is independently depolarized when exposed to light having a first wavelength and wherein the second light activated protein is independently depolarized when exposed to light having a second wavelength;
- wherein the first or the second light activated protein is a chimeric protein derived from VChR1 from Volvox carteri and ChR1 from Chlamydomonas reinhardti, wherein the protein comprises the amino acid sequence of VChR1 having at least the first and second transmembrane helices replaced by the first and second transmembrane helices of ChR1.
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 derived from VChR1 from Volvox carteri and ChR1 from Chlamydomonas reinhardti, wherein the protein comprises the amino acid sequence of VChR1 having at least the first and second transmembrane helices replaced by the first and second transmembrane helices of ChR1;
- (b) measuring an excitatory post synaptic potential (EPSP) in response to selectively depolarizing the excitatory neuron comprising a first light-activated protein or measuring an inhibitory post synaptic current (IPSC) in response to selectively depolarizing an inhibitory neuron comprising a 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.
Dependencies are inferred from the claim text.