Light-sensitive ion-passing molecules
Disclosed are polynucleotides and methods for expressing light activated proteins in animal cells and altering an action potential of the cells by optical stimulation. The disclosure also provides animal cells and non-human animals comprising cells expressing the light-activated proteins.
1. An animal cell comprising a light-activated protein expressed on the cell membrane, wherein the protein comprises, in order from amino terminus (N-terminus) to carboxyl terminus (C-terminus):
a) a core amino acid sequence at least 95% identical to the sequence shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, or SEQ ID NO:4;
b) an endoplasmic reticulum (ER) export signal; and
c) a membrane trafficking signal.
2. The animal cell of claim 1 , wherein the ER export signal comprises the amino acid sequence FCEYENEV (SEQ ID NO:12).
3. The animal cell of claim 1 , wherein the membrane trafficking signal comprises the amino acid sequence KSRITSEGEYIPLDQIDINV (SEQ ID NO:11).
4. The animal cell of claim 1 , wherein the animal cell is a neuronal cell, a muscle cell, or a stem cell.
5. The animal cell of claim 1 , further comprising a second light-activated protein expressed on the cell membrane.
6. The animal cell of claim 5 , wherein the second light-activated protein is a protein capable of mediating a depolarizing current in the cell when the cell is illuminated with light.
7. The animal cell of claim 5 , wherein the second light-activated protein is selected from the group consisting of a VChR1, a DChR, a SFO, and a ChR2.
8. A population of cells comprising the cell of claim 1 .
9. A method of using the animal cell of claim 1 , the method comprising activating the light-activated protein with light.
10. The method of claim 9 , wherein the light-activated protein comprises an amino acid sequence having at least 95% amino acid sequence identity to the amino acid sequence set forth in SEQ ID N0:3, wherein the light has a wavelength of between about 400 and 740 nm.
11. The method of claim 10 , wherein the light has a wavelength of 570 nm.
12. The method of claim 10 , wherein the cell further comprise a second light-activated protein expressed on the cell membrane, and wherein the method comprises activating the light-activated proteins at their respective wavelengths.
13. The method of claim 12 , wherein the second light-activated protein is a protein capable of mediating a depolarizing current in the cell when the cell is illuminated with light.
14. The method of claim 12 , wherein the second light-activated protein is selected from the group consisting of a VChR1, a DChR, a SFO, and a ChR2.