The KCTD3 Knockout SK-HEP-1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population in which the KCTD3 gene is disrupted in the SK-HEP-1 human hepatocellular carcinoma cell line. This tool provides a loss-of-function model for investigating KCTD3 in the context of liver cancer, particularly its roles in the ubiquitin-proteasome system and actin cytoskeleton regulation. The polyclonal format reflects a heterogeneous pool of edited cells, offering a robust representation of knockout effects without clonal selection.
SK-HEP-1 cells were originally isolated from the ascitic fluid of a patient with liver adenocarcinoma and are widely adopted as a model for hepatocellular carcinoma. This cell line recapitulates key malignant features and is frequently employed to study hepatocarcinogenesis, cell migration, and therapeutic responses. Its well-annotated genomic and phenotypic profiles make it an ideal host for gene editing studies.
KCTD3 encodes a substrate adaptor of the Cullin-3 (CUL3)-RING E3 ubiquitin ligase complex. It mediates the ubiquitination and proteasome-dependent degradation of the small GTPases RhoA and Rac1, thereby modulating actin cytoskeleton dynamics, cell polarity, and migration. Through its interaction with CUL3 and the RING protein RBX1, KCTD3 links the ubiquitin-proteasome system to Rho GTPase signaling, controlling the stability of key regulators of the actin network.
Disruption of KCTD3 in SK-HEP-1 hepatocellular carcinoma cells is expected to impair the degradation of RhoA and Rac1, leading to altered actin filament organization and migratory behavior. This enables researchers to dissect the contribution of KCTD3 to processes relevant to tumor invasion and metastasis. The model serves as a platform to uncover KCTD3-dependent mechanisms that drive hepatocellular carcinoma progression.
These polyclonal knockout cells support a broad spectrum of applications, including functional genomics, mechanistic studies of ubiquitin-mediated signaling, and phenotypic assays for cell migration (e.g., wound healing and Transwell). They are well-suited for target validation experiments assessing the impact of KCTD3 loss on proliferation, motility, or drug sensitivity. Standard readouts include Western blotting for KCTD3 and its targets, quantitative RT-PCR, RNA sequencing, immunofluorescence for actin cytoskeleton visualization, RhoA activation assays, and co-immunoprecipitation to confirm disruption of CUL3 interactions. For further information or to order, please contact Ascent Research.
