Light-activated cation channel and uses thereof

摘要

The present invention provides compositions and methods for light-activated cation channel proteins and their uses within cell membranes and subcellular regions. The invention provides for proteins, nucleic acids, vectors and methods for genetically targeted expression of light-activated cation channels to specific cells or defined cell populations. In particular the invention provides millisecond-timescale temporal control of cation channels using moderate light intensities in cells, cell lines, transgenic animals, and humans. The invention provides for optically generating electrical spikes in nerve cells and other excitable cells useful for driving neuronal networks, drug screening, and therapy.

主权项

1. A method of predicting potential ion channel modulating properties of a drug, the method comprising:
a) labeling a neuronal cell with a voltage sensitive dye, wherein the neuronal cell stably expresses a light-activated cation channel protein and an ion channel of interest, wherein the light-activated cation channel protein comprises an amino acid sequence having at least 75% amino acid sequence identity to ChR2; b) exposing the neuronal cell to light, wherein the light is delivered at a frequency of at least 20 Hz; c) monitoring the voltage sensitive dye to determine a first response in said cell; d) exposing said dye-labeled neuronal cell to a candidate drug; e) after exposing said cell to the candidate drug, further exposing said cell to light; f) monitoring the voltage sensitive dye to determine a second response in said cell; and g) comparing said first and second responses to identify a potential ion channel modulating property of said candidate drug, wherein a second response that is higher than the first response indicates that the candidate drug modulates the ion channel.