Channelrhodopsins for optical control of cells
The invention, in some aspects relates to compositions and methods for altering cell activity and function and the introduction and use of light-activated ion channels.
1. A method of altering ion conductivity of a membrane, the method comprising, expressing in a membrane in a cell a light-activated ion channel polypeptide encoded in a recombinant nucleic acid vector comprising a heterologous nucleic acid molecule insert encoding a light-activated ion channel amino acid polypeptide sequence, the amino acid sequence having at least 90% amino acid identity to amino acids 86-320 as set forth in SEQ ID NO: 2 and at least 95% identity to the amino acids 321-350 as set forth as SEQ ID NO: 2 and contacting the expressed light-activated ion channel polypeptide with a light that activates the light-activated ion channel polypeptide and alters the ion conductivity of the membrane.
2. The method of claim 1 , wherein the cell is a mammalian cell.
3. The method of claim 1 , wherein the cell is an excitable cell.
4. The method of claim 1 , wherein altering the ion conductivity of the membrane comprises depolarizing the membrane.
5. A method of assessing the effect of a candidate compound on ion conductivity of a membrane, the method comprising,
(a) expressing in a membrane in a cell a light-activated ion channel polypeptide encoded in a recombinant nucleic acid vector comprising a heterologous nucleic acid molecule insert encoding a light-activated ion channel amino acid polypeptide sequence, the amino acid sequence having at least 90% amino acid identity to amino acids 86-320 as set forth in SEQ ID NO: 2 and at least 95% identity to the amino acids 321-350 as set forth as SEQ ID NO: 2;
(b) contacting the membrane comprising the light-activated ion channel polypeptide with light under conditions suitable for altering ion conductivity of the membrane;
(c) contacting the membrane with a candidate compound at one or both of before and after contacting with the light; and
(d) identifying the presence or absence of a change in ion conductivity of the membrane contacted with the light and the candidate compound compared to an ion conductivity in a control cell membrane comprising the expressed light-activated ion channel polypeptide and contacted with the light and not contacted with the candidate compound; wherein a change in the ion conductivity in the membrane compared to the ion conductivity in the control membrane indicates an effect of the candidate compound on the ion conductivity of the membrane.
6. The method of claim 5 , wherein the cell is a mammalian cell.
7. The method of claim 5 , wherein the cell is an excitable cell.
8. The method of claim 5 , wherein the effect of the candidate compound on the ion conductivity of the membrane is depolarization of the membrane.
9. The method of claim 1 , wherein the expressed light-activated ion channel polypeptide is part of a heterologous fusion protein.
10. The method of claim 5 , wherein the expressed light-activated ion channel polypeptide is part of a heterologous fusion protein.