ANKRD28 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population derived from the HT29 human colorectal adenocarcinoma line, featuring targeted disruption of the ANKRD28 gene. This knockout model enables investigation of ANKRD28-dependent processes without the need for single-cell cloning, maintaining the heterogeneity of the parental population while abrogating ANKRD28 protein expression. The polyclonal format offers a robust system for functional genomics studies, where gene disruption is achieved across the bulk population, providing a loss-of-function context for downstream assays.
The host cell line, HT29, is an epithelial cell line established from a primary colon adenocarcinoma of a 44-year-old Caucasian female. HT29 cells are extensively characterized in colorectal cancer research, exhibiting adherent growth and retaining key features of intestinal epithelial cells. They are commonly employed to study tumor biology, drug responses, and signal transduction pathways relevant to colon carcinogenesis. The use of HT29 as the parental line provides a clinically relevant background for investigating the role of ANKRD28 in colorectal cancer progression.
ANKRD28 encodes a scaffold protein containing ankyrin repeat domains that functions as a regulatory subunit for protein phosphatase 1 (PP1). It directly interacts with PP1 catalytic subunits??PPP1CA, PPP1CB, and PPP1CC??to form holoenzyme complexes that control substrate specificity. By targeting PP1 activity to cytoskeletal elements such as myosin light chain and actin-associated proteins, ANKRD28 plays a critical role in cytoskeletal organization and remodeling. This dephosphorylation signaling axis intersects with cell cycle regulation and motility pathways. Although upstream regulators of ANKRD28 remain poorly defined, its downstream effects are mediated through PP1 substrate dephosphorylation, impacting actomyosin contractility and cellular architecture.
In the context of HT29 colorectal adenocarcinoma cells, disruption of ANKRD28 expression is expected to perturb PP1-mediated dephosphorylation events, potentially altering cytoskeletal integrity and signaling outputs. Because ANKRD28 scaffolds PP1 to substrates involved in cell adhesion and migration, its knockout may influence the metastatic behavior and proliferative capacity of these cancer cells. This model thus provides a valuable system for dissecting how ANKRD28-dependent PP1 regulation contributes to colorectal tumorigenesis and for evaluating whether ANKRD28 represents a modulatory node in colon cancer progression.
Researchers can apply ANKRD28 Knockout HT29 Polyclonal Cells to a broad range of experimental workflows. The cells are well-suited for western blotting and RT-qPCR to confirm loss of ANKRD28 expression, as well as immunofluorescence and co-immunoprecipitation to assess PP1 complex integrity and localization. Functional studies may include cell proliferation assays, migration assays, and phospho-signaling analyses to probe the impact on downstream pathways. Additionally, the knockout model can be integrated into drug sensitivity screens and flow cytometry-based phenotyping to explore ANKRD28 as a modulator of therapeutic response in colorectal cancer. For additional details or to inquire about this product, please contact Ascent Research.
