KBTBD8 Knockout HCT 116 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the KBTBD8 gene in the human HCT 116 colorectal carcinoma cell line. This heterogeneous polyclonal pool contains diverse KBTBD8 gene disruptions, providing a loss-of-function system suitable for functional studies while minimizing clone-specific artifacts. The knockout is achieved using CRISPR/Cas9-mediated gene disruption without implying monoclonality or specific editing outcomes.
The HCT 116 cell line, derived from a primary human colon carcinoma, is an epithelial-like, DNA mismatch repair-proficient model widely used in colorectal cancer research. Its well-defined genetics and reproducible growth characteristics make it an ideal host for gene-editing experiments, enabling clean dissection of gene function in a relevant tumor context.
KBTBD8 encodes a substrate-recognition adaptor for the Cullin3-RBX1 E3 ubiquitin ligase complex. It specifically targets Inhibitor of DNA-binding proteins ID1, ID2, and ID3, as well as TBX6, for polyubiquitination and proteasomal degradation. Through this mechanism, KBTBD8 modulates TGF-beta/BMP signaling, influencing downstream effectors such as SMAD1/5/8. Upstream regulators including BMP4 and WNT3A control KBTBD8 expression, placing it within a network regulating neural crest specification and mesoderm development. KBTBD8 interacts directly with CUL3 and RBX1 to form the active ligase complex.
In HCT 116 cells, KBTBD8 knockout likely stabilizes Id proteins, altering TGF-beta/BMP signaling and potentially affecting cell proliferation, differentiation, and migration. Given the established role of Id proteins in cancer stemness and therapy resistance, this knockout model provides a platform to investigate KBTBD8??s contributions to colorectal cancer progression. The genetic stability of HCT 116 cells allows reliable assessment of KBTBD8-dependent ubiquitination dynamics.
This polyclonal knockout population is suitable for functional assays including western blotting and RT-qPCR for target validation, ubiquitination and co-immunoprecipitation studies to probe CUL3 complex interactions, and proteasome inhibition experiments to confirm substrate stabilization. Phenotypic analyses such as cell proliferation, migration, and invasion assays are recommended to assess the impact on tumorigenic properties. Additional applications encompass RNA-seq transcriptomic profiling, luciferase reporter assays for TGF-beta/BMP pathway activity, and drug target validation. For further information or to discuss custom gene-editing services, please contact Ascent Research.
