BIRC2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 colorectal adenocarcinoma cell line. This product provides a heterogeneous pool of cells with targeted disruption of the BIRC2 gene, enabling loss-of-function studies in an intestinal epithelial cancer model suitable for apoptosis and NF-??B signaling research.
The HT29 cell line was established from a primary colon adenocarcinoma of a 44-year-old female and is widely used as a model for colorectal cancer biology. These adherent epithelial cells retain key malignant features, including dysregulated proliferation and survival pathways, and offer a genetically stable background for CRISPR-based gene disruption, making them a standard tool for investigating oncogenic mechanisms and drug responses.
BIRC2 (cIAP1) is an E3 ubiquitin ligase that inhibits apoptosis by binding and suppressing caspase-3 and caspase-7. It also serves as a critical regulator of canonical NF-??B signaling downstream of TNF-??. Upon TNFR1 activation, BIRC2 is recruited to receptor complexes with TRADD, TRAF2, and RIP1, where it catalyzes K63-linked ubiquitination of RIP1, promoting IKK complex assembly and subsequent NF-??B nuclear translocation. This activity is counteracted by SMAC/DIABLO and modulated by interactions with XIAP. Through these mechanisms, BIRC2 drives expression of pro-survival and proliferative genes at the intersection of apoptosis suppression and inflammatory signaling.
Endogenous BIRC2 in HT29 cells supports malignant phenotypes by blocking apoptosis and sustaining NF-??B-dependent transcription of survival and inflammatory genes. BIRC2 disruption in this background sensitizes cells to death receptor agonists and may impair growth factor-induced NF-??B activation, underscoring colorectal cancer??s reliance on IAP-mediated cytoprotection. These knockout cells therefore provide a relevant model to study apoptosis threshold modulation, cytokine signaling rewiring, and therapeutic targeting using SMAC mimetics or other IAP antagonists.
These polyclonal knockout cells enable diverse experimental applications. Western blotting for BIRC2, cleaved caspases, and phosphorylated I??B?? confirms target disruption and pathway effects. Viability assays (MTT/resazurin) and Annexin V/PI flow cytometry quantify apoptosis and drug sensitivity. Caspase activity and NF-??B luciferase reporter assays directly measure signaling outputs, while RT-qPCR assesses transcriptional changes in downstream targets. Co-immunoprecipitation studies reveal altered protein interactions in the TNFR1 complex. For technical details, please contact Ascent Research.
