KLHL12 Knockout HCT 116 Polyclonal Cells provide a CRISPR/Cas9-edited knockout cell population specifically targeting the KLHL12 gene in the human HCT 116 colorectal carcinoma cell line. This polyclonal format yields a heterogeneous loss-of-function model, avoiding clonal biases while enabling robust investigation of KLHL12-dependent processes. The product is designed for researchers studying ubiquitin-mediated regulation, COPII vesicle trafficking, and Wnt signaling in a physiologically relevant cancer background.
HCT 116 is a widely characterized colorectal carcinoma epithelial line harboring an activating KRAS G13D mutation, homozygous MLH1 deficiency, and a microsatellite instability?high (MSI?H) phenotype, together with a near?diploid karyotype. These features render HCT 116 a valuable model for studying oncogenic KRAS signaling, DNA mismatch repair defects, and MSI?H tumor biology. Its adherent growth and epithelial differentiation make it suitable for mechanistic studies of tumor cell behavior and drug response, providing a consistent platform for gene disruption experiments.
KLHL12 serves as a substrate adaptor for the CUL3?RBX1 E3 ubiquitin ligase, directing the ubiquitination of SEC31A to regulate COPII coat assembly and efficient ER?to?Golgi transport of large cargo proteins, including procollagen. Concurrently, KLHL12 interacts with Dishevelled (DVL1, DVL2, DVL3) proteins to promote their degradation, thereby dampening Wnt/???catenin signaling downstream of WNT3A activation. Through these dual roles, KLHL12 integrates collagen secretory pathway activity with Wnt signal transduction. The adaptor also partners with ALG2 and PEF1 to modulate cargo selection, linking ubiquitin?proteasome function to COPII?dependent secretion. Disrupting KLHL12 enables dissection of these interconnected mechanisms controlling extracellular matrix production and signal propagation.
In HCT 116 cells, loss of KLHL12 can shed light on how altered collagen secretion and Wnt pathway dysregulation contribute to colorectal cancer pathogenesis. The KRAS G13D mutation drives persistent proliferative and pro?survival signals, while MLH1 deficiency and MSI?H status create a mutagenic environment; KLHL12 knockout may modify collagen deposition in the tumor microenvironment and influence ???catenin?dependent transcription. This model therefore enables the study of matrix remodeling and signaling crosstalk under conditions mimicking aggressive colorectal carcinoma, with potential implications for understanding fibrosis?associated cancer progression and drug resistance.
Typical applications include COPII trafficking assays via immunofluorescence for SEC31A and collagen, collagen secretion analysis by Western blotting, and Wnt target gene quantification by RT?qPCR. Co?immunoprecipitation of CUL3 complexes paired with in?cell ubiquitination assays facilitates interrogation of E3 ligase activity. Drug sensitivity profiling with MEK inhibitors, migration assays, and transcriptomic analyses such as RNA?seq can be performed to assess therapeutic vulnerabilities and signaling perturbations. For technical inquiries, please contact Ascent Research.
