633 / 2019-04-27 21:14:12

CRISPR/Cas9-mediated knockout of IP3Rs diminishes store-operated Ca2+ entry of HEK293 cells.

IP3Rs, calcium channel

全体主题 > Signal Transduction

Ca2+ release from intracellular Ca2+ store and Ca2+ entry across plasma membrane (PM), are sources of Ca2+ signals. As a main intracellular Ca2+ store with high concentration of Ca2+ levels, endoplasmic reticulum (ER) play an essential role in Ca2+ signaling. Proper Ca2+ concentration within ER is necessary for ER function and cell development. In this study, we first generated IP3Rs triple and double knockout HEK cell lines (IP3Rs-TKO and IP3Rs-DKO) using CRISPR/Cas9 genome-editing technology. Using these engineered cells with clean genetic background, we demonstrate that IP3Rs-TKO impairs ER calcium homeostasis in HEK cells, which lead to reduction of ER Ca2+ content, resulting multimerization of a small amount of STIM1 and subsequent coupling of STIM1 and Orai1, ultimately cause minor constitutive Ca2+ entry. In addition, IP3Rs-TKO also diminish SOCE through promoting Orai1 protein degradation, that may be a negative regulation mechanism response constitutive Ca2+ entry. That provides a train of thought for the study of plant ion channel.