Optogenetic probes for measuring membrane potential
AssigneesPresident and Fellows of Harvard College, The Governors of the University of Alberta
InventorsAdam E. Cohen, Daniel Hochbaum, Peng Zou, Samouil Leon Farhi, Robert Earl Campbell, Yongxin Zhao, Daniel Jed Harrison
Life of this patent
Estimating the term…
Abstract
Provided herein are variants of an archaerhodopsin useful for application such as optical measurement of membrane potential. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, cells comprising the polynucleotides, and cells comprising the polypeptides; and methods of using the variants.
Claims21
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises at least one substitution mutation selected from the group consisting of P60S, T80S, D95H, D95Q, D106H, and F161V, wherein the position of the at least one substitution mutation in the amino acid sequence corresponds to SEQ ID NO:1.
A polynucleotide comprising a nucleic acid sequence encoding a variant polypeptide having at least 80% identity to SEQ ID NO:
- 1, wherein the variant polypeptide comprises at least one substitution mutation selected from the group consisting of P60S, T80S, D95H, D95Q, D106H, and F161V, wherein the position of the at least one substitution mutation in the amino acid sequence corresponds to SEQ ID NO:
- 1.
A polynucleotide comprising a nucleic acid sequence of SEQ ID NO:
- 5 or SEQ ID NO:
- 6.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a first substitution mutation at position 95, and at least one second substitution mutation selected from the group consisting of P60S, T80S, D106H, and F161V, wherein the positions of the substitution mutations in the amino acid sequence corresponds to SEQ ID NO:1.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a substitution mutation at position 95, a D106H mutation, and at least one substitution mutation selected from the group consisting of P60S, T80S, and F161V, wherein the positions of the substitution mutations in the amino acid sequence corresponds to SEQ ID NO:1.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a substitution mutation at position 95 of D to Q, C, or N, and
- at least one substitution mutation selected from the group consisting of P60S, T80S, and F161V;
- wherein the positions of the substitution mutations in the amino acid sequence corresponds to SEQ ID NO:1.
A polynucleotide encoding a polypeptide comprising an amino acid sequence of SEQ ID NO:
- 2 or SEQ ID NO:
- 3.
A fusion protein comprising an amino acid sequence of SEQ ID NO:
- 2 or SEQ ID NO:
- 3 and an amino acid sequence of a second fluorescent protein.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a D to H or D to Q substitution mutation at position 95, wherein the position of the substitution mutation in the amino acid sequence corresponds to SEQ ID NO:
- 1.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a P to S substitution mutation at position 60, wherein the position of the substitution mutation in the amino acid sequence corresponds to SEQ ID NO:
- 1.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a T to S substitution mutation at position 80, wherein the position of the substitution mutation in the amino acid sequence corresponds to SEQ ID NO:
- 1.
A polypeptide comprising a variant having at least 80% identity to SEQ ID NO:
- 1, wherein the amino acid sequence of the variant comprises a D to H substitution mutation at position 106, wherein the position of the substitution mutation in the amino acid sequence corresponds to SEQ ID NO:
- 1.
Dependencies are inferred from the claim text.