This product is a CRISPR/Cas9-edited polyclonal knockout cell population derived from Huh-7 cells, in which the DTNA gene has been disrupted to create a loss-of-function model of alpha-dystrobrevin. The polyclonal format provides a mixed population of cells with heterogeneous gene edits, facilitating population-level analyses without single-cell cloning. This knockout model is designed for researchers investigating the roles of the dystrophin-glycoprotein complex in hepatocellular carcinoma biology.
The Huh-7 cell line is a widely used human hepatocellular carcinoma model, originally established from a well-differentiated liver tumor. These epithelial cells retain key liver functions and are commonly employed in studies of liver metabolism, hepatocellular carcinoma progression, and hepatitis C virus replication. Their tumorigenic properties, including anchorage-dependent growth and invasive potential, make them particularly suitable for examining mechanisms of cell adhesion, migration, and signal transduction in liver cancer.
DTNA encodes alpha-dystrobrevin, a critical scaffolding component of the dystrophin-associated glycoprotein complex (DGC) that bridges the intracellular actin cytoskeleton and the extracellular matrix. Alpha-dystrobrevin forms direct interactions with dystrophin, utrophin, alpha- and beta-syntrophin, and the sarcoglycan complex, and it serves to anchor neuronal nitric oxide synthase (nNOS) at the plasma membrane. Upstream, DTNA expression is regulated by the muscle-specific transcription factors MyoD and MEF2, and its activity is modulated by mechanical stretch. Through these interactions, alpha-dystrobrevin facilitates DGC assembly, membrane stability, and signal integration via nitric oxide and MAPK pathways, ultimately impacting cellular mechanotransduction.
Disruption of DTNA in the Huh-7 hepatocellular carcinoma model provides a powerful tool to investigate the contributions of the dystrophin-glycoprotein complex to liver cancer biology. Loss of alpha-dystrobrevin may alter cell?Cmatrix adhesion, migration, and invasion??processes that are critical for tumor metastasis. This model enables the study of DGC function in a non-muscle epithelial context, where its roles are not well defined, and allows for the examination of how DTNA knockout impacts the composition and stability of DGC-associated complexes, potentially revealing new therapeutic targets for hepatocellular carcinoma.
This polyclonal knockout cell population is ideal for a range of experimental workflows. Researchers can assess DTNA and DGC protein expression by western blotting, visualize actin cytoskeleton and dystrophin localization via immunofluorescence, and measure cell adhesion and migration/invasion capabilities. Co-immunoprecipitation of dystrophin and syntrophin can delineate complex formation in the absence of alpha-dystrobrevin, while RNA sequencing can identify downstream transcriptional changes. Moreover, apoptosis assays and drug sensitivity studies can evaluate compounds aimed at restoring DGC function or targeting related signaling pathways. For more information or to discuss custom projects, please contact Ascent Research.
