The CASP9 Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human NCI-H1975 non-small cell lung carcinoma (NSCLC) line. This heterogeneous pool carries a targeted disruption of the CASP9 gene introduced by CRISPR/Cas9-mediated gene editing, yielding a loss-of-function model for apoptosis research without clonal selection bias.
The NCI-H1975 host cell line is a well-characterized human lung adenocarcinoma model bearing EGFR L858R and T790M mutations, along with wild-type TP53. These genetic features underlie its widespread use in studying EGFR tyrosine kinase inhibitor (TKI) resistance and the apoptotic evasion that often accompanies treatment failure.
CASP9 initiates the intrinsic apoptosis pathway. Upon mitochondrial outer membrane permeabilization, cytochrome c (CYCS) is released and binds APAF1, forming the apoptosome that activates procaspase-9. Active CASP9 then cleaves executioner caspases CASP3 and CASP7, which in turn target substrates such as PARP1 and DFFA. Upstream, BCL2 family proteins including BAX regulate mitochondrial integrity, while TP53 promotes apoptosis transcriptionally. Negative regulation is mediated by XIAP and BIRC5 through direct caspase inhibition.
In NCI-H1975 cells, CASP9 is central to apoptosis commitment, connecting mitochondrial stress to executioner caspase activation. The EGFR T790M mutation often confers apoptosis resistance, limiting TKI efficacy. Disruption of CASP9 enables dissection of apoptosis bypass mechanisms and the identification of strategies to re-engage cell death in resistant NSCLC.
This product supports diverse applications: monitoring apoptosome assembly via co-immunoprecipitation, detecting cytochrome c release, measuring caspase-3/7 activity, and performing western blots for cleaved CASP9. Annexin V flow cytometry quantifies apoptosis, while cell viability assays with EGFR inhibitors assess therapeutic responses. The polyclonal population is also suited for pro-apoptotic compound screening and genetic complementation studies. For additional information, please contact Ascent Research.
