The KLHL13 Knockout HCT 116 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HCT 116 human colorectal carcinoma cell line. This heterogeneous pool of cells carries a diverse array of loss-of-function mutations in the KLHL13 gene introduced by CRISPR/Cas9-mediated gene disruption. The polyclonal format minimizes clone-specific biases and provides a genetically diverse background, enabling robust functional genomics studies. This knockout population serves as a versatile tool for dissecting KLHL13-dependent pathways without the need for monoclonal isolation, facilitating reproducible and translationally relevant experimental outcomes.
The HCT 116 host cell line is a human colorectal carcinoma epithelial cell line widely employed in cancer research. These cells are characterized by their adherent growth, stable karyotype, and well-defined signaling networks, making them a standard model for colorectal tumorigenesis and drug response investigations. HCT 116 cells are particularly suitable for cell cycle studies due to their predictable mitotic progression and intact DNA damage checkpoints. Their amenability to CRISPR/Cas9 gene editing and compatibility with numerous downstream assays further support their use in knockout-based functional analyses.
KLHL13 acts as a substrate-specific adaptor for the Cullin3?CRBX1 E3 ubiquitin ligase complex, which catalyzes the ubiquitination and degradation of mitotic regulators including PLK1 and Aurora B. This targeted proteolysis is essential for proper mitotic progression and cytokinesis. KLHL13 operates within the ubiquitin?Cproteasome system, interacting with Cullin3, RBX1, and ubiquitin-conjugating enzymes to control substrate stability. Disruption of KLHL13 leads to accumulation of its targets, causing mitotic defects and failed cell division.
In the context of HCT 116 colorectal carcinoma cells, loss of KLHL13 disrupts the controlled degradation of PLK1 and Aurora B, causing impaired mitotic progression and cytokinesis failure. This model captures the consequences of defective ubiquitin-mediated proteolysis, a hallmark of many colorectal cancers where dysregulation of E3 ligase adaptors contributes to genomic instability and therapeutic resistance. The polyclonal nature of the knockout population allows for the assessment of phenotype robustness across a mixed genetic landscape, enhancing its utility for translational research. By studying KLHL13 loss, researchers can identify vulnerabilities specific to colorectal cancer and explore mechanisms of drug sensitivity.
This KLHL13 knockout polyclonal cell product is ideally suited for a range of biomedical applications, including investigation of ubiquitin-dependent cell cycle regulation, functional studies of the Cullin3?CRBX1 ligase complex, and drug sensitivity profiling. Representative assays include western blotting to monitor PLK1 and Aurora B protein levels, ubiquitination assays to assess substrate modification, flow cytometry-based cell cycle analysis, and immunofluorescence for mitotic markers. Additionally, colony formation and drug dose-response assays enable quantification of long-term proliferative capacity and chemosensitivity. The model also supports synthetic lethality screens and combinatorial drug testing in a colorectal cancer background. For further technical details or to discuss customized applications, please contact Ascent Research.
