The DOCK2 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal population of HEK293T cells with targeted disruption of the DOCK2 gene. This loss-of-function model enables dissection of DOCK2-dependent cellular processes without the need for transient silencing or repeated transfections. As a polyclonal knockout pool, it offers a genetically heterogeneous background that can be leveraged for stable expression studies, functional screens, and pathway interrogation where clonal variation is minimized through population-level analysis.
The parental HEK293T cell line is a widely utilized human embryonic kidney epithelial model, transformed with SV40 large T-antigen to promote high-level protein expression and efficient viral production. These adherent cells exhibit robust growth characteristics and high transfection efficiency, making them an ideal chassis for genetic manipulation. While originally derived from kidney epithelium, HEK293T cells express neuronal lineage markers, offering a unique platform for studying signaling pathways that bridge epithelial and neuronal cell biology.
DOCK2 (dedicator of cytokinesis 2) encodes a guanine nucleotide exchange factor (GEF) that specifically activates the small GTPases RAC1 and CDC42, master regulators of actin cytoskeleton dynamics. Mechanistically, DOCK2 functions downstream of chemokine receptors such as CXCR4 and CCR7, integrins, and Src family kinases, integrating signals from cytokines like IL-2 and IL-7. Upon activation, DOCK2, in complex with ELMO1, facilitates GDP-GTP exchange on RAC1. Active RAC1 then stimulates downstream effectors including PAK1, the WAVE complex, and LIMK, leading to cofilin inactivation and Arp2/3-mediated actin polymerization. This signaling axis drives membrane protrusion, cell migration, and adhesion, processes essential for immune cell activation and metastatic dissemination.
Although DOCK2 is predominantly characterized in hematopoietic cells, its role in actin remodeling and migration is conserved across cell types. In HEK293T epithelial cells, DOCK2 knockout provides a reductionist model to study actin-dependent phenomena such as cell spreading, migration, and adhesion in a non-hematopoietic context. This model is particularly valuable for dissecting the cell-autonomous functions of DOCK2 in cytoskeletal dynamics without the complexity of immune receptor signaling, and for investigating its potential contributions to epithelial-mesenchymal transition and cancer cell metastasis.
This polyclonal knockout cell pool is suitable for a variety of downstream applications. Knockout efficiency can be confirmed by western blotting, Sanger sequencing, or RT-qPCR. Functional consequences can be assessed by RAC1 activity assays, transwell migration assays, and phalloidin staining to visualize F-actin organization. Additional applications include immunofluorescence for cytoskeletal architecture, cell adhesion assays, and drug screening for compounds that modulate DOCK2-dependent pathways. Researchers studying immunodeficiencies, autoimmune diseases, or anticancer therapies can employ these cells for mechanistic studies and high-content screening. For further information, please contact Ascent Research.
