The BIRC2 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population derived from HEK293T, providing a loss-of-function model for the anti-apoptotic E3 ligase cIAP1. This polyclonal format yields a heterogeneous gene-disrupted pool ideal for robust phenotype analysis, transient complementation experiments, and pooled functional genomics screens, without clonal selection artifacts.
HEK293T is a female human embryonic kidney cell line immortalized with sheared adenovirus 5 DNA and stably expressing the SV40 large T antigen. This background confers exceptionally high transfection efficiency and supports episomal replication of plasmids containing the SV40 origin, making it a preferred host for lentivirus production, protein overexpression, and signaling pathway dissection. Its well-characterized signaling networks and rapid growth further solidify its utility for generating CRISPR-mediated knockout models.
The BIRC2 gene encodes cIAP1, a pivotal inhibitor of apoptosis and E3 ubiquitin ligase that regulates canonical and non-canonical NF-??B signaling. Downstream of TNFR, cIAP1 ubiquitinates RIP1 to activate the IKK complex and NF-??B p65, while simultaneously targeting NIK for constitutive ubiquitination and proteasomal degradation in the alternative pathway. Pro-apoptotic SMAC/DIABLO antagonizes cIAP1, releasing caspase-3, -7, and -9 to execute cell death. Upstream stimuli include TNF-??, IL-1??, and LPS, and cIAP1 interacts with TRAF1, TRAF2, and XIAP to fine-tune inflammatory and survival responses.
In the HEK293T context, BIRC2 knockout profoundly sensitizes cells to TNF-???Cinduced apoptosis and attenuates NF-??B activation, enabling precise dissection of cIAP1??s role in RIP1-dependent necroptosis and NIK-mediated non-canonical signaling. The high transfectability of these cells facilitates complementation with domain mutants or inhibitor studies, and supports generation of double knockouts with interacting partners such as XIAP or TRAF2, enhancing the model??s versatility for probing apoptotic network dynamics.
These polyclonal knockout cells are applied in apoptosis assays with caspase-3/7 activity measurements, cell viability analyses under death receptor stimulation, and NF-??B luciferase reporter assays. Co-immunoprecipitation and ubiquitination assays elucidate cIAP1 substrate interactions, while RT-qPCR and Western blotting confirm downstream target regulation. The model supports drug resistance mechanism studies for SMAC mimetics, proteasome inhibitors, and novel ubiquitin-proteasome system modulators. For additional information or to place an order, please contact Ascent Research.
