The DTNA Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout population derived from the HT29 human colorectal adenocarcinoma cell line, designed to disrupt the DTNA gene encoding alpha-dystrobrevin. This heterogeneous loss-of-function model avoids clonal artifacts and enables robust functional interrogation of alpha-dystrobrevin-dependent processes in a genetically tractable epithelial system. The polyclonal composition is ideal for population-level assays of cell adhesion, signaling dynamics, and differentiation.
The HT29 cell line, isolated from a primary human colorectal adenocarcinoma (Dukes?? stage B), is a well-established intestinal epithelial model. These adherent cells possess a hypertriploid karyotype and can differentiate into enterocyte-like cells upon treatment with inducing agents. Widely used in gastrointestinal research, HT29 cells recapitulate key aspects of colonocyte biology, including polarization and barrier function, providing a physiologically relevant host for studying DTNA in colorectal cancer and normal colon physiology.
Alpha-dystrobrevin (DTNA) scaffolds the dystrophin-associated protein complex (DAPC) at the cell membrane, linking dystroglycan and integrin adhesion receptors to the actin cytoskeleton. It directly associates with dystrophin, syntrophins, and dystroglycan, recruiting signaling effectors including nNOS, GRB2, and dysbindin. Knockout of DTNA in HT29 cells destabilizes DAPC integrity, impairing focal adhesion kinase (FAK) and Src family kinase activation downstream of integrins and growth factor receptors such as EGFR. Consequently, downstream MAPK/ERK and PI3K-AKT pathways are attenuated, and nNOS-mediated signaling is disrupted, collectively altering actin remodeling, focal adhesion turnover, and transcriptional responses (e.g., MMPs, cytokines).
In HT29 colorectal adenocarcinoma cells, DTNA knockout is expected to compromise cell-matrix adhesion and mechanotransduction, influencing epithelial homeostasis, anoikis sensitivity, and differentiation. The DAPC has been implicated in tumor suppression, and this polyclonal model enables dissection of dystrobrevin-dependent pathways that may govern cancer cell migration, invasion, and survival. By avoiding muscle-specific complexities, the HT29 system offers a simplified carcinoma platform to study DAPC function in a gastrointestinal malignancy context.
These polyclonal knockout cells support diverse applications, including western blotting and RT-qPCR for validating target disruption, immunofluorescence for DAPC localization, and co-immunoprecipitation to probe protein interactions. Functional assays such as transwell migration/invasion, cell adhesion, and anoikis assays elucidate alpha-dystrobrevin??s role in epithelial behavior, while phospho-protein arrays and flow cytometry enable pathway profiling and apoptosis/proliferation analysis. The cells are also suited for compound screening to restore DAPC function or target downstream effectors. For additional information or to place an order, contact Ascent Research.
