The KBTBD4 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, engineered to disrupt the KBTBD4 gene. This loss-of-function model enables investigation of KBTBD4-dependent processes in a cancer-relevant context. The polyclonal format provides a heterogeneous population of knockout cells, suitable for pooled functional studies.
HT29 cells originated from a primary colorectal adenocarcinoma of a 44-year-old female and exhibit an aneuploid karyotype with well-characterized mutations in tumor suppressor genes APC and TP53, among other cancer-associated alterations. This cell line is widely employed as a model for intestinal epithelial differentiation and colorectal cancer progression, offering a physiologically relevant backdrop for studying oncogenic signaling and therapeutic responses.
KBTBD4 encodes a substrate adaptor for the CUL3-RBX1 E3 ubiquitin ligase complex. It recognizes specific protein substrates, facilitating their ubiquitination by an E2 conjugating enzyme and subsequent proteasomal degradation. Within this ubiquitin-proteasome pathway, KBTBD4 interacts directly with CUL3 and RBX1, while the broader complex includes ubiquitin, proteasome subunits, and E2 conjugating enzymes. Although upstream regulators of KBTBD4 remain largely uncharacterized, it may be transcriptionally modulated by stress-responsive factors. Downstream, KBTBD4-mediated ubiquitination controls the stability of target proteins, thereby modulating cellular signaling networks.
In HT29 colorectal adenocarcinoma cells, disruption of KBTBD4 is expected to impair the CUL3-RBX1-mediated ubiquitination of its substrates, potentially leading to dysregulated protein turnover. This perturbation may alter pathways governing cell proliferation, differentiation, and stress adaptation, providing insight into the role of ubiquitin-dependent proteolysis in colorectal cancer biology. The model thus serves as a valuable tool for dissecting how aberrations in the ubiquitin-proteasome system contribute to tumorigenesis.
This polyclonal knockout cell population is suited for a range of functional applications, including mechanistic studies of the ubiquitin-proteasome system, investigation of protein degradation dynamics, and cancer cell biology research. Researchers can employ these cells in assays such as Western blotting to assess protein abundance, RT-qPCR and RNA-seq for transcriptomic profiling, ubiquitination assays to monitor substrate modification, as well as cell viability, colony formation, and migration assays to evaluate phenotypic consequences. Additionally, the cells facilitate drug target validation and functional genomics screening. For further details, please contact Ascent Research.
