IBTK Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population generated from the HT29 human colorectal adenocarcinoma cell line, designed to disrupt the IBTK gene. This product provides a heterogeneous cell pool with IBTK gene disruption, enabling loss-of-function studies without requiring single-cell cloning. The polyclonal format offers a robust model for investigating IBTK-dependent biology while maintaining genetic diversity that more closely reflects population-level responses.
The HT29 cell line originates from a primary colorectal adenocarcinoma of a 44-year-old female and is widely employed as an intestinal epithelial model for studying barrier function, cancer biology, and drug screening. HT29 cells retain many characteristics of colonic epithelium, including the ability to differentiate into enterocyte-like cells, making them a versatile system for colorectal cancer research.
IBTK encodes an inhibitor of Bruton’s tyrosine kinase (BTK), functioning as a substrate adaptor for a Cullin 3 (CUL3)-based ubiquitin ligase complex. Upon BCR activation and downstream NF-??B signaling, IBTK interacts with BTK and promotes its ubiquitination and proteasomal degradation. This process suppresses BTK-mediated phosphorylation of PLC??2 and subsequent NF-??B activation, thereby reducing transcription of anti-apoptotic target genes such as BCL2 and BCL-XL. By negatively regulating BCR-like signaling, IBTK facilitates apoptosis induction and acts as a potential tumor suppressor.
In the HT29 colorectal cancer context, loss of IBTK is predicted to enhance survival signaling by disrupting its negative regulation of BTK. Although BTK is classically associated with B-cell receptor signaling, emerging evidence suggests that BTK expression and BCR-like signaling components play roles in colorectal tumorigenesis. This knockout model enables dissection of IBTK??s tumor-suppressive functions and investigation of how aberrant ubiquitination-dependent regulation of BTK contributes to colorectal cancer progression and resistance to apoptosis.
Researchers can employ this polyclonal knockout model to elucidate IBTK function in colorectal cancer, explore non-canonical roles of BTK inhibitors in solid tumors, and study cross-talk between ubiquitination machinery and survival signaling. Typical downstream assays include Western blotting for BTK, phospho-BTK, and NF-??B targets; RT-qPCR for BCL2 and c-MYC; apoptosis evaluation via Annexin V/PI staining; cell proliferation using MTT or BrdU; ubiquitination co-immunoprecipitation; flow cytometric cell cycle analysis; colony formation assays; and transcriptome profiling by RNA-seq. For further details, please contact Ascent Research.
