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US 2025/0154221 A1Published application
Voltage indicators for two-photon imaging
OwnerAhmed Abdelfattah, Rosario Valenti +1inventors
InventorsAhmed Abdelfattah, Rosario Valenti, Eric R. Schreiter
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Abstract
A voltage indicator is described, including a voltage-sensitive rhodopsin domain having mutations to replace canonical counterion charge residues with neutral residues, and a capture protein that binds a fluorescent element, wherein the capture protein is provided together with the voltage-sensitive rhodopsin domain in a fusion protein.
Claims20
A voltage indicator, comprising:
- a voltage-sensitive rhodopsin domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:
- 1-26, including a mutation to replace a charged canonical amino acid residue with a neutral residue at one, two, or three residues within a pump or channel core motif of the rhodopsin domain;
- and
- a capture protein that covalently or noncovalently binds a fluorescent element that is a fluorescent protein, or a fluorescent dye;
- wherein the capture protein is provided together with the voltage-sensitive rhodopsin domain in a fusion protein.
2 more elements
A nucleotide molecule, comprising a nucleotide encoding an amino acid sequence selected from the group consisting of SEQ ID NO:
- 1-26, including a mutation to replace a charged canonical amino acid residue with a neutral residue at a residue within a pump or channel core motif of the rhodopsin domain.
A method of measuring voltage, the method comprising:
- (i) delivering to a cell a nucleotide encoding an amino acid molecule comprising
- a voltage-sensitive rhodopsin domain comprising an amino acid sequence selected from the group consisting of SEQ ID NO:
- 1-26, including a mutation to replace a charged canonical amino acid residue with a neutral residue at a residue within a pump or channel core motif of the rhodopsin domain;
- and
- a capture protein that covalently or noncovalently binds a fluorescent element that is a fluorescent protein, or a fluorescent dye;
- wherein the capture protein is provided together with the voltage-sensitive rhodopsin domain in a fusion protein;
- and
- (ii) delivering to the cell a fluorescent element that is a fluorescent protein or a fluorescent dye, wherein
- when the fluorescent element is a fluorescent protein, it is provided together with the capture protein and the voltage-sensitive rhodopsin domain in a fusion protein;
- and
- when the fluorescent element is a fluorescent dye, it is attached to a ligand of the capture protein;
- and
- (iii) determining changes in fluorescence of the fluorescent element, as an measurement of voltage changes in the cell, wherein an increase in membrane potential lead to an increase in fluorescence.
10 more elements
Dependencies are inferred from the claim text.