Light-sensitive ion-passing molecules
AssigneeThe Board of Trustees of the Leland Stanford Junior University
InventorsKarl Deisseroth, Feng Zhang, Viviana Gradinaru
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Abstract
The invention provides polynucleotides and methods for expressing light-activated proteins in animal cells and altering an action potential of the cells by optical stimulation. The invention also provides animal cells and non-human animals comprising cells expressing the light-activated proteins.
Claims27
A method of identifying transsynaptic connection between neuronal cells in an animal or a tissue, comprising:
- a) administering a first viral vector encoding a Cre recombinase fused to a transcellular tracer protein to neuronal cells in region A of the animal or tissue, wherein the transcellular tracer protein is wheat germ agglutinin (WGA) or tetanus toxin-fragment C (TTC), wherein the first viral vector is selected from the group consisting of an adenoassociated virus vector, a herpes simplex virus vector, and a lentiviral vector;
- b) administering a second viral vector encoding a light-activated protein to neuronal cells in region B of the animal or tissue, wherein the expression of the light-activated protein depends on the presence of the Cre recombinase, wherein the light-activated protein comprises an amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:3, SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:4, wherein the second viral vector is selected from the group consisting of an adenoassociated virus vector, a herpes simplex virus vector, and a lentiviral vector;
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- c) identifying neuronal cells expressing the light-activated protein in region B, wherein the expression of the light-activated protein in the neuronal cells indicates that these cells are in transsynaptic connection with the cells in region A.
A method of generating optical control of targeted neuronal cells in an animal or tissue, comprising:
- a) administering a first viral vector expressing a Cre recombinase fused to a transcellular tracer protein to region A of the animal or tissue, wherein the transcellular tracer protein is wheat germ agglutinin (WGA) or tetanus toxin-fragment C (TTC), wherein the first viral vector is selected from the group consisting of an adenoassociated virus vector, a herpes simplex virus vector, and a lentiviral vector;
- b) administering a second viral vector encoding a light-activated protein to region B of the animal or tissue, wherein the expression of the light-activated protein depends on the presence of the Cre recombinase, wherein the neuronal cells in region A and in region B are in transsynaptic connection, and wherein the light-activated protein comprises an amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:3, SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:4, wherein the second viral vector is selected from the group consisting of an adenoassociated virus vector, a herpes simplex virus vector, and a lentiviral vector;
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- c) controlling action potential of a neuronal cell in region B with light that activates the light-activated protein.
A method of controlling action potential of a neuron in an animal, comprising activating a light-activated protein in the neuron with light to generate action potential change, wherein expression of the light-activated protein in the neuron is generated by:
- a) administering a first viral vector expressing a Cre recombinase fused to a transcellular tracer protein to region A of the animal, wherein the transcellular tracer protein is wheat germ agglutinin (WGA) or tetanus toxin-fragment C (TTC), wherein the first viral vector is selected from the group consisting of an adenoassociated virus vector, a herpes simplex virus vector, and a lentiviral vector;
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- b) administering a second viral vector encoding a light-activated protein to region B of the animal which contains the neuron, wherein the expression of the light-activated protein depends on the presence of the Cre recombinase, wherein the neurons in region A and region B are in transsynaptic connection, wherein the light-activated protein comprises an amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:3, SEQ ID NO:1, SEQ ID NO:2, or SEQ ID NO:4, wherein the second viral vector is selected from the group consisting of an adenoassociated virus vector, a herpes simplex virus vector, and a lentiviral vector.
Dependencies are inferred from the claim text.