The ITPR1 Knockout NCI-H1975 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal human cell population with targeted disruption of the ITPR1 gene, establishing a loss-of-function model for inositol 1,4,5-trisphosphate receptor type 1. Derived from the NCI-H1975 lung adenocarcinoma cell line, this polyclonal knockout pool enables functional studies without clonal selection artifacts. CRISPR/Cas9-mediated gene disruption allows investigation of ITPR1-dependent calcium signaling and downstream cellular processes in a cancer-relevant context.
The parental NCI-H1975 cell line is a well-characterized human lung adenocarcinoma epithelial model harboring EGFR L858R and T790M mutations, which drive constitutive kinase activity and confer resistance to first- and third-generation EGFR tyrosine kinase inhibitors (TKIs). This NSCLC adenocarcinoma model recapitulates key clinical resistance mechanisms and is widely employed for mechanistic studies and drug evaluation. The epithelial adherent morphology supports diverse in vitro assay formats.
ITPR1 encodes the type 1 IP3 receptor, an ER-resident calcium channel. It is activated by IP3 generated downstream of G??q-coupled GPCRs or receptor tyrosine kinases such as EGFR via phospholipase C. Calcium release through ITPR1 activates calmodulin, calcineurin, NFAT, CaMKII, and PKC, regulating transcription, proliferation, and apoptosis. ITPR1 interacts with calmodulin, FKBP12, IRBIT, and Bcl-2; Bcl-2 binding inhibits calcium release and suppresses apoptosis. Additionally, ITPR1-mediated mitochondrial calcium uptake facilitates cytochrome c release, linking ER calcium to cell death execution.
In NCI-H1975 cells, EGFR signaling through PLC?? generates IP3, engaging ITPR1-dependent calcium mobilization. Knockout of ITPR1 may uncouple EGFR activity from calcium-mediated survival and proliferative pathways, potentially resensitizing cells to EGFR TKIs. The ITPR1?CBcl-2 interaction suggests that loss of ITPR1 could lower the apoptotic threshold. Moreover, calcium oscillations governed by ITPR1 impact migration and invasion, making this model valuable for studying metastatic behavior in the context of TKI resistance.
These polyclonal knockout cells are suited for investigating calcium signaling in EGFR-mutant lung cancer, exploring mechanisms of TKI resistance, screening for agents targeting ER calcium release, and studying apoptosis via the Bcl-2?CITPR1 axis. Common assays include Western blot, RT-qPCR, Fluo-4 AM calcium imaging, flow cytometry with calcium indicators, Annexin V apoptosis assays, and transwell migration/invasion tests. Osimertinib sensitivity profiling can also be performed. For further information or technical support, contact Ascent Research.
