The DOCK2 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the HT29 human colorectal adenocarcinoma cell line, featuring targeted disruption of the DOCK2 gene. This product provides a heterogeneous pool of edited cells, each carrying distinct genomic modifications within the DOCK2 locus, enabling loss-of-function studies without the bottleneck of single-cell cloning. The polyclonal format preserves population-level diversity while ensuring robust abrogation of DOCK2 protein expression. By eliminating DOCK2 function, researchers can dissect its role in Rac GTPase signaling and associated cellular processes in a model system that retains the intrinsic characteristics of the parental HT29 line.
HT29 is a well-characterized human colorectal adenocarcinoma epithelial cell line that serves as a standard intestinal epithelial model. These cells exhibit typical epithelial features such as polarized morphology, tight junction formation, and mucin production, making them valuable for studying intestinal barrier function, nutrient transport, and epithelial homeostasis. The tumorigenic nature of HT29 cells also renders them a widely employed platform for colorectal cancer research, including analyses of cell proliferation, differentiation, and oncogenic signaling. As a human-derived line, HT29 overcomes species-specific limitations, offering a clinically relevant context for exploring gene function in colorectal epithelial biology and pathology.
DOCK2 (Dedicator of Cytokinesis 2) functions as a guanine nucleotide exchange factor (GEF) for Rac1 and Rac2, small GTPases that orchestrate actin cytoskeleton remodeling. In the canonical signaling cascade, chemokine receptors such as CXCR4 and CCR7, upon ligand binding, activate G??i-coupled heterotrimeric G proteins, leading to PI3K-mediated production of phosphatidylinositol-3,4,5-trisphosphate (PIP3). PIP3 recruits DOCK2, often in complex with the adaptor ELMO1, to the plasma membrane, where it catalyzes GDP/GTP exchange on Rac. Active Rac then engages downstream effectors including PAK1, the WAVE regulatory complex, and Arp2/3, culminating in actin polymerization and lamellipodia formation. DOCK2 also interacts with CRKL, VAV1, and other signaling nodes, linking it to broader networks of integrin and immune receptor pathways. Knockout of DOCK2 thus ablates a critical nexus for chemokine-induced cell migration and cytoskeletal dynamics.
In the HT29 colorectal adenocarcinoma context, disruption of DOCK2 profoundly impacts Rac-dependent processes that are integral to cancer cell behavior. Since HT29 cells retain epithelial characteristics and migratory capacity, loss of DOCK2 is anticipated to impair actin-dependent motility, invasion, and potentially chemokine-directed migration, mimicking aspects of metastatic suppression. This model therefore provides a tractable system to investigate how DOCK2-mediated actin remodeling contributes to colorectal cancer progression. Moreover, DOCK2 deficiency has been associated with primary immunodeficiency and inflammatory bowel disease, positioning this knockout model as a tool for exploring how intestinal epithelial DOCK2 influences mucosal immune homeostasis and epithelial barrier integrity under pathophysiological conditions.
Researchers can employ the DOCK2 Knockout HT29 Polyclonal Cells in a broad range of experimental applications. Functional rescue via chemokine stimulation (e.g., CXCL12/CXCR4) can be compared between wild-type and knockout cells to assess migration and invasion defects. Representative assays include cell migration and invasion assays, Rac activation pull-downs, phospho-PAK western blotting, phalloidin-based immunofluorescence for F-actin visualization, and flow cytometric evaluation of chemokine receptor surface expression. Transcriptional profiling by RNA-seq or targeted RT-qPCR can reveal DOCK2-dependent gene signatures. This knockout model is also suitable for drug screening campaigns aimed at identifying immunomodulatory or anti-metastatic compounds. For further technical details or customized reagent requests, please contact Ascent Research.
