The IKBIP Knockout NCI-H1975 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the NCI-H1975 human lung adenocarcinoma cell line. This product provides a heterogeneous pool of cells with targeted disruption of the IKBIP gene, serving as a loss-of-function model to investigate IKBIP??s tumor-suppressive roles. The polyclonal format eliminates the need for single-cell cloning and facilitates robust, population-level analyses in signaling and apoptosis research.
The NCI-H1975 host cell line is an epithelial lung adenocarcinoma model derived from a female patient, harboring EGFR L858R and T790M mutations. These mutations are oncogenic drivers and key determinants of sensitivity and resistance to EGFR tyrosine kinase inhibitors (TKIs) such as erlotinib and osimertinib. Widely used in NSCLC research, NCI-H1975 cells provide a clinically relevant platform for studying apoptosis, NF-kappa-B signaling, and TKI resistance mechanisms.
IKBIP functions as a pro-apoptotic protein by directly interacting with IKK-beta (IKBKB) and TRAF2, forming complexes that suppress canonical NF-kappa-B signaling. This interaction prevents phosphorylation and degradation of NFKBIA (I??B??), blocking RELA (p65) nuclear translocation and transcription of anti-apoptotic genes such as BCL2L1 and XIAP. Simultaneously, IKBIP promotes JNK phosphorylation, cytochrome c release, and caspase-3/7 (CASP3) activation. Upstream, IKBIP is regulated by TP53, ER stress (involving HSPA5 and DDIT3), and DNA damage signals, integrating stress inputs to drive apoptosis through dual NF-kappa-B inhibition and JNK-caspase activation.
Disruption of IKBIP in NCI-H1975 cells is expected to derepress NF-kappa-B signaling, enhancing RELA transcriptional activity and upregulating anti-apoptotic factors, thereby conferring resistance to ER stress- and DNA damage-induced apoptosis. In the context of EGFR-mutant lung adenocarcinoma, this knockout may promote TKI resistance, making it a valuable model for investigating apoptosis evasion and identifying synthetic lethal vulnerabilities. The model also facilitates studies on how loss of a pro-apoptotic tumor suppressor intersects with oncogenic EGFR signaling.
This knockout cell product enables a broad array of applications, including NF-kappa-B signaling studies, apoptosis resistance analysis, and EGFR TKI resistance research. Investigators can employ Western blotting for IKBIP, IKBKB, phospho-RELA, and cleaved CASP3; RT-qPCR for NF-kappa-B target genes; Annexin V flow cytometry; caspase-3/7 activity assays; NF-kappa-B luciferase reporter assays; and cell viability or drug sensitivity testing with erlotinib or osimertinib. Co-immunoprecipitation of IKBKB and functional screening for synthetic lethality are also supported. For additional details, please contact Ascent Research.
