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US 12,105,122 B2Granted
Voltage indicators
AssigneeHoward Hughes Medical Institute
InventorsEric R. Schreiter, Ahmed Abdelfattah
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Abstract
A voltage indicator includes a polypeptide sequence comprising a voltage-sensitive opsin domain and a capture protein domain arranged and disposed to capture a fluorescent dye ligand. When the fluorescent dye ligand is captured and the voltage indicator is bound to a cell membrane, an increase in voltage across the cell membrane causes an increase in fluorescent emission.
Claims20
A method of measuring voltage, the method comprising
- (a) providing a voltage indicator, comprising:
- (i) a voltage-sensitive microbial rhodopsin domain comprising a polypeptide selected from a group consisting of QuarsAr1, QuarsAr2, Ace2N, and combinations thereof including one, two, three, or four amino acid mutations relative to a wild type polypeptide sequence;
- and
- (ii) a capture protein that covalently or noncovalently binds a fluorescent dye ligand that is a fluorescent protein;
- or
- a fluorescent dye, wherein the capture protein is provided together with the voltage-sensitive microbial rhodopsin domain in a fusion protein-;
- (b) contacting the voltage indicator and the fluorescent dye ligand with a cell, and
- (c) determining changes in fluorescence of the fluorescent dye ligand when the fluorescent dye ligand is captured by the voltage indicator.
5 more elements
A voltage indicator, comprising:
- a voltage-sensitive microbial rhodopsin domain comprising a polypeptide selected from the group consisting of QuarsAr1, QuarsAr2, Ace2N, and combinations thereof including one, two, three, or four amino acid mutations relative to a wild type polypeptide sequence;
- and
- a capture protein that covalently or noncovalently binds a fluorescent dye ligand that is
- a fluorescent protein;
- or
- a fluorescent dye, wherein the capture protein is provided together with the voltage-sensitive microbial rhodopsin domain in a fusion protein.
3 more elements
Dependencies are inferred from the claim text.