Granted Patent
B2
US 12,331,089 · App. 17/668,766 · Granted Jun 17, 2025
Channelrhodopsins for optical control of cells
Inventors:
Nathan Klapoetke (Ashburn, VA); Brian Yichiun Chow (Cherry Hill, NJ); Edward Boyden (Chestnut Hill, MA); Gane Ka-Shu Wong (Edmonton, CA); Yongku Peter Cho (Vernon, CT)
C07K14/405A61K41/0023A61N5/062C12N13/00G01N33/48728G01N33/5041G01N33/5058A61K38/00A61N2005/0661A61N2005/0662
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Abstract
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.
Claims (15)
1. A method of producing spikes in a neuron, the method comprising:
a) introducing into a neuron a nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of a light-activated ion channel polypeptide, 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, to generate a modified neuron that expresses the light-activated ion channel polypeptide; and
b) contacting the light-activated ion channel polypeptide in the modified neuron with a light wherein the contacting activates the light-activated ion channel polypeptide, alters the ion conductivity of the neuron membrane, and induces spikes in the modified neuron.
2. The method of claim 1 , wherein the light-activated ion channel polypeptide is Chrimson ChR88 and has the amino acid sequence of SEQ ID NO: 2.
3. The method of claim 1 , wherein the light-activated ion channel polypeptide is ChR88 K176R and has the amino acid sequence of SEQ ID NO: 5.
4. The method of claim 1 , wherein the light-activated ion channel polypeptide is part of a heterologous fusion protein.
5. The method of claim 1 , wherein the neuron is a visual system cell.
6. The method of claim 1 , wherein the light has a wavelength ranging from 365 nm to 700 nm.
7. The method of claim 1 , wherein the light has a wavelength ranging from 530 nm to 640 nm.
8. The method of claim 3 , wherein the light has a wavelength of 590 nm +/−15 nm.
9. The method of claim 1 , wherein the light has pulse widths ranging from 1 ms to 5 ms.
10. The method of claim 1 , wherein the light has powers ranging from 0.1 mW/mm 2 to 100 mW/mm 2 .
11. The method of claim 3 , wherein the light-activated ion channel polypeptide generates spikes from 1 to 10 mW mm −2 .
12. The method of claim 1 , wherein the nucleic acid is a viral vector.
13. The method of claim 12 , wherein the viral vector is an adeno-associated virus.
Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Feb 17, 2022
From: WONG, GANE KA-SHU
To: THE GOVERNORS OF THE UNIVERSITY OF ALBERTA
Reel/Frame 059030/0515 →
ASSIGNMENT OF ASSIGNOR'S INTEREST
Recorded Feb 17, 2022
From: KLAPOETKE, NATHAN; CHOW, BRIAN YICHIUN; BOYDEN, EDWARD; CHO, YONGKU PETER
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 059030/0550 →
Continuity (5)
Continuation
14357635
Provisional Application
61559076
· Nov 12, 2011
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