The ACTN1 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population, derived from the HT29 human colorectal adenocarcinoma cell line, engineered for targeted disruption of the ACTN1 gene encoding alpha-actinin-1. This heterogeneous knockout pool provides a loss-of-function model for studying ACTN1-dependent processes without clonal selection bias, enabling robust phenotypic analysis of cytoskeletal and adhesion dynamics.
HT29 cells are an extensively characterized epithelial model of colorectal cancer, originally isolated from a primary adenocarcinoma. They retain features of intestinal epithelial differentiation and are widely used in cancer biology, drug response, and adhesion studies. Their adherent, epithelial phenotype renders them suitable for investigating cell-matrix and cell-cell interactions governed by the actin cytoskeleton and focal adhesion dynamics.
Alpha-actinin-1 is an actin crosslinking protein that bundles F-actin filaments and anchors them to focal adhesions and adherens junctions through interactions with vinculin, paxillin, integrin beta, and alpha- and beta-catenin. ACTN1 is activated downstream of integrin engagement, FAK/Src signaling, and mechanical tension, and it regulates actin stress fiber formation, focal adhesion assembly, and cell migration, in part by modulating RhoA pathway activity. This positions ACTN1 as a structural and signaling hub coupling the actin cytoskeleton to both cell-matrix and cell-cell adhesion complexes.
In HT29 colorectal cancer cells, ACTN1 contributes to invasive and migratory behavior, with upregulation linked to metastatic progression. Disruption of ACTN1 in this polyclonal population allows investigation of how loss of this crosslinker alters adhesion strength, cytoskeletal tension, and migratory velocity. This model is particularly relevant for dissecting mechanisms of metastasis where dynamic actin remodeling and focal adhesion turnover are critical.
Applications include western blot and RT-qPCR confirmation of knockout and downstream pathway analysis, immunofluorescence for actin and focal adhesion visualization, functional assays such as transwell migration, invasion, and adhesion assays, and co-immunoprecipitation to probe protein interactions. The model also supports time-lapse motility imaging and drug screening targeting ACTN1-dependent metastasis mechanisms. For further product details and technical support, please contact Ascent Research.
