The IKBKB Knockout Ca Ski Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the Ca Ski human cervical carcinoma cell line, designed for loss-of-function studies of the IKBKB gene (encoding IKK??). This product consists of a heterogeneous pool of knockout cells generated via CRISPR/Cas9-mediated gene disruption, enabling investigation of IKK??-dependent signaling without clonal artifacts.
The Ca Ski cell line originates from a cervical epidermoid carcinoma metastasis and is a well-established model of HPV-16-positive cervical cancer. These cells harbor integrated HPV-16 genomes, and the viral oncoproteins E6 and E7 inactivate the tumor suppressors p53 and Rb, respectively, thereby recapitulating key oncogenic features of cervical carcinogenesis.
IKBKB encodes IKK??, the catalytic subunit of the IKK complex, which also includes IKK?? (CHUK) and the regulatory subunit NEMO (IKBKG). IKK?? is central to canonical NF-??B signaling, phosphorylating I??B?? (NFKBIA) and I??B?? on conserved serine residues (e.g., Ser32/36 of I??B??), leading to their ubiquitination and proteasomal degradation. This releases NF-??B dimers??principally p65 (RELA)/p50 (NFKB1)??which translocate to the nucleus and transactivate genes encoding pro-inflammatory cytokines (e.g., IL-6, IL-8, TNF), anti-apoptotic factors (e.g., Bcl-xL, XIAP, cIAPs), and cell cycle regulators (e.g., cyclin D1, c-Myc). IKK?? is activated by upstream stimuli including TNF-??, IL-1??, LPS, and antigen receptors, via adaptor molecules such as TRAF2, RIP1, and the TAK1/TAB kinase complex.
In Ca Ski cells, the constitutive activation of NF-??B contributes to the malignant phenotype driven by HPV oncoproteins, promoting inflammation, survival, and proliferation. Knockout of IKBKB in this background abrogates canonical NF-??B responses, providing a powerful tool to dissect the interplay between viral transformation and host innate immunity. This model enables the study of how IKK??-dependent signaling modulates cervical cancer cell behavior, including cytokine production, apoptosis resistance, and invasive potential, in the context of HPV-16 positivity.
Typical applications include analyzing NF-??B signaling dynamics following stimulation with TNF-?? or IL-1?? using western blotting for phospho-I??B??, NF-??B reporter assays, and ELISA for secreted IL-6 and IL-8. The cells are suitable for screening IKK?? inhibitors, examining apoptosis induction (Annexin V/PI staining), and assessing cell migration and invasion. Co-immunoprecipitation can verify IKK complex integrity, while immunofluorescence monitors p65 nuclear translocation. For detailed technical specifications or batch-specific validation data, please contact Ascent Research.
