Agonist-Induced Ca²⁺ Signaling in HEK-293-Derived Cells Expressing a Single IP₃ Receptor Isoform.

ABSTRACT: In mammals, three genes encode IP₃ receptors (IP₃Rs), which are involved in agonist-induced Ca²⁺ signaling in cells of apparently all types. Using the CRISPR/Cas9 approach for disruption of two out of three IP₃R genes in HEK-293 cells, we generated three monoclonal cell lines, IP3R1-HEK, IP3R2-HEK, and IP3R3-HEK, with the single functional isoform, IP₃R1, IP₃R2, and IP₃R3, respectively. All engineered cells responded to ACh with Ca²⁺ transients in an "all-or-nothing" manner, suggesting that each IP₃R isotype was capable of mediating CICR. The sensitivity of cells to ACh strongly correlated with the affinity of IP₃ binding to an IP₃R isoform they expressed. Based on a mat

SUBMITTER: Kochkina EN 

PROVIDER: S-EPMC11011116 | biostudies-literature | 2024 Mar

REPOSITORIES: biostudies-literature