The KLHL7 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population of HT29 human colorectal adenocarcinoma cells with targeted disruption of the KLHL7 gene. This heterogeneous pool of knockout cells, generated by CRISPR/Cas9-mediated genome editing, provides a loss-of-function model for investigating KLHL7-dependent mechanisms in a cancer-relevant background. The polyclonal format avoids clonal bias, enabling robust analysis of gene function across a diverse cellular context.
The HT29 cell line, derived from a colon adenocarcinoma of a 44-year-old female, is a widely used epithelial model for colorectal cancer research. These cells exhibit adherent growth and retain key oncogenic features, including dysregulated proliferation and impaired cell cycle control, making them particularly suitable for studying the interplay between ubiquitin-mediated proteolysis and tumor cell biology. The integration of KLHL7 knockout into this background allows direct assessment of its role in mitotic regulation within colon adenocarcinoma.
KLHL7 functions as a substrate adaptor for the CUL3-RBX1 ubiquitin ligase complex, mediating the K48-linked polyubiquitination and proteasomal degradation of specific target proteins. Central to its function is the recognition and targeting of the mitotic kinase Aurora B (AURKB) for destruction, a process governed by upstream transcriptional regulators and cell cycle signals. Disruption of KLHL7 abolishes substrate recruitment, leading to Aurora B accumulation and subsequent mitotic defects, including abnormal spindle formation and chromosome segregation errors, thereby compromising genomic stability.
In HT29 colorectal cancer cells, KLHL7 knockout polyclonal cells enable detailed dissection of the ubiquitin-proteasome system’s role in maintaining mitotic fidelity. Accumulation of mitotic substrates upon KLHL7 loss can drive chromosomal instability, a hallmark of colorectal cancer progression. This model facilitates investigation of how impaired degradation of Aurora B and other regulators contributes to unchecked proliferation and tumor cell survival, offering insights into potential therapeutic vulnerabilities associated with ubiquitin pathway dysfunction.
Typical applications include western blotting to assess AURKB protein stability, flow cytometry for cell cycle analysis, and immunofluorescence to visualize mitotic spindle abnormalities. Co-immunoprecipitation experiments can probe CUL3 complex assembly and substrate interactions, while proliferation assays and mitotic index scoring quantify cell division defects. These polyclonal knockout cells also support drug target screening for agents that modulate mitotic catastrophe or exploit synthetic lethality in colorectal cancer. For further information, please contact Ascent Research.
