Genetically encoded calcium indicator polypeptides and nucleic acids encoding same
Provided herein are nucleic acid sequences and polypeptides encoding a genetically encoded calcium indicator (GECI). Also provided are vectors and cells comprising the nucleic acid sequences and/or polypeptides. Kits comprising the nucleic acid sequences, polypeptides, vectors, cells and combinations thereof are also provided. Also provided herein are methods of screening for G-protein coupled receptor (GPCR) agonists and antagonists and methods of monitoring neural activity using the GECIs.
1. An isolated nucleic acid sequence encoding a genetically encoded calcium indicator (GECI), wherein the GECI comprises SEQ ID NO: 2.
2. A vector comprising the isolated GECI-encoding nucleic acid sequence of claim 1 .
3. An isolated cell comprising the isolated GECI-encoding nucleic acid sequence of claim 1 .
4. The isolated cell of claim 3 , wherein the GECI-encoding nucleic acid sequence is located in the genome of the cell.
5. An isolated polypeptide comprising a GECI, wherein the GECI comprises SEQ ID NO: 2.
6. An isolated cell comprising the isolated polypeptide of claim 5 .
7. The isolated cell of claim 3 , wherein the cell further comprises a G-protein coupled receptor (GPCR).
8. The isolated cell of claim 7 , wherein the cell comprises a nucleic acid sequence encoding the GPCR.
9. The isolated cell of claim 8 , wherein the nucleic acid sequence encoding the GPCR is located on a vector or in the genome of the cell.
10. The isolated cell of claim 7 , wherein the GPCR is selected from the group consisting of alpha-1 adrenegic receptors (I1-AR), urotensin (UT) receptors, 5-HT2, 5-HT6 and 5-HT7 serotonin receptors, hypocretic (orexin) receptors, histamine H1 receptors, bradykinin receptors, bombesin receptors, purinergic receptors, acetycholine receptors, mGluR5 glutamate receptors, vasopressin receptors, angiotensin receptors, cholecystokinin receptors, endothelin receptors, ghrelin receptors, melatonin receptors, neurotensin receptors, platelet-activating factor receptors, prolactin releasing peptide receptors, GABA-B receptors, histamine receptors, and glutamate receptors.
11. The isolated cell of claim 3 , wherein the cell is a Hela cell or a human embryonic kidney (HEK) cell.
12. The isolated cell of claim 11 , wherein the HEK cell is a HEK293 cell.
13. The isolated cell of claim 6 , wherein the cell further comprises a G-protein coupled receptor (GPCR).
14. The isolated cell of claim 13 , wherein the cell comprises a nucleic acid sequence encoding the GPCR.
15. The isolated cell of claim 14 , wherein the nucleic acid sequence encoding the GPCR is located on a vector or in the genome of the cell.
16. The isolated cell of claim 13 , wherein the GPCR is selected from the group consisting of alpha-1 adrenegic receptors (I1-AR), urotensin (UT) receptors, 5-HT2, 5-HT6 and 5-HT7 serotonin receptors, hypocretic (orexin) receptors, histamine H1 receptors, bradykinin receptors, bombesin receptors, purinergic receptors, acetycholine receptors, mGluR5 glutamate receptors, vasopressin receptors, angiotensin receptors, cholecystokinin receptors, endothelin receptors, ghrelin receptors, melatonin receptors, neurotensin receptors, platelet-activating factor receptors, prolactin releasing peptide receptors, GABA-B receptors, histamine receptors, and glutamate receptors.
17. The isolated cell of claim 6 , wherein the cell is a Hela cell or a human embryonic kidney (HEK) cell.
18. The isolated cell of claim 17 , wherein the HEK cell is a HEK293 cell.