The DIAPH3 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal human cell population with targeted disruption of DIAPH3. This model is produced from the HT29 parental line using CRISPR/Cas9 to introduce gene disruption, generating a heterogeneous pool of knockout cells. The polyclonal format allows for functional studies of DIAPH3 loss-of-function without clonal isolation, providing a practical tool for population-level analyses.
The HT29 cell line is a widely used human colorectal adenocarcinoma model with epithelial morphology, established from a primary tumor of a 44-year-old female. HT29 cells retain characteristics of intestinal epithelial cells, including the ability to differentiate under appropriate conditions, and are commonly employed in studies of colon cancer biology, epithelial barrier function, and drug response. This well-characterized line provides a physiologically relevant background for investigating the role of DIAPH3 in colorectal cancer progression.
DIAPH3 encodes a member of the formin family of actin nucleation factors that promotes the assembly of unbranched actin filaments. Activated downstream of RhoA GTPase and ROCK signaling, DIAPH3 interacts with profilin and G-actin to drive actin polymerization, and associates with APC and EB1 to coordinate microtubule dynamics. Through these interactions, DIAPH3 regulates stress fiber formation, focal adhesion turnover, cell migration, and cytokinesis. The protein serves as a key effector in Rho GTPase-mediated cytoskeletal reorganization, linking extracellular cues from integrin and EGF receptor signaling to actin remodeling.
In HT29 colorectal adenocarcinoma cells, DIAPH3 disruption impairs the formation of contractile actin stress fibers and destabilizes focal adhesions, leading to reduced cell motility and compromised invasive capacity. Given that DIAPH3 is implicated in cancer metastasis and its expression is dysregulated in colorectal tumors, this knockout model enables dissection of the molecular mechanisms by which formin-driven actin dynamics contribute to epithelial tumor progression. The HT29 background offers a tractable system to examine how loss of DIAPH3 affects colon cancer cell behavior, including adhesion, migration, and responses to chemotherapeutic agents.
This polyclonal knockout cell population is optimally suited for a range of functional assays, including scratch wound healing and Transwell invasion assays to assess cell migration and invasion, immunofluorescence staining of F-actin, vinculin, and paxillin to visualize cytoskeletal structures and focal adhesions, and RhoA activation assays to probe upstream signaling. Additional applications encompass cell adhesion, proliferation, and apoptosis analyses, as well as RT-qPCR for epithelial-mesenchymal transition (EMT) markers and co-immunoprecipitation to examine DIAPH3 protein interactions. For more information or to inquire about this product, please contact Ascent Research.
