Low affinity red fluorescent indicators for imaging ca2+ in excitable and nonexcitable cells
AssigneesThe Governors of the University of Alberta, The Chancellor, Masters and Scholars of the University of Oxford
InventorsYu-Fen Chang, Jiahui Wu, Matthew J. Daniels, Robert E. CAMPBELL
Life of this patent
Estimating the term…
Abstract
The present disclosure relates to genetically encoded low affinity, fluorescent Ca2+ indicators, which may be targeted to endoplasmic reticulum, the sarcoplasmic reticulum and/or the mitochondria. It also relates to polynucleotides, vectors and host cells which encode or include such low affinity Ca2+ indicators, and methods of detecting Ca2+ levels in a cell using such indicators.
Claims25
A method of detecting changes in Ca2+ levels in a cell, the method comprising:
- a. obtaining a sample comprising cells engineered to express one or more low affinity Ca2+ indicator selected from the group consisting of:
- SEQ ID Nos 4, 6, 8, 10, 12, 14, 16 and 18, or a polypeptide that has at least 90% sequence identity to any one of the foregoing, which is fluorescent and has an affinity for Ca2+ with a Kd of greater than 20 but excluding SEQ ID NO. 2;
- b. exposing the cells to excitation light;
- and
- c. detecting changes in ER, SR and/or mitochondria Ca2+ levels by visualizing or imaging the cells.
A low affinity fluorescent Ca2+ polypeptide selected from the group consisting of:
- SEQ ID Nos 4, 6, 8, 10, 12, 14, 16 and 18, or a polypeptide that has at least 90% sequence identity to any one of the foregoing, which is fluorescent and has an affinity for Ca2+ with a Kd of greater than 20 μM, but excluding SEQ ID NO. 2.
(canceled)
(canceled)
(canceled)
(canceled)
Dependencies are inferred from the claim text.