DPYSL2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human embryonic kidney cell line HEK293T, carrying a targeted disruption of the DPYSL2 gene. This loss-of-function model enables researchers to study the cellular consequences of DPYSL2 ablation without introducing specific predefined mutations, as the polyclonal pool contains a diverse array of CRISPR-induced edits that cumulatively eliminate functional protein expression across the population.
The parental HEK293T cell line is a widely utilized tool in biomedical research, originating from HEK293 cells and transformed with sheared adenovirus type 5 DNA. These cells constitutively express the SV40 large T antigen, which permits episomal replication of plasmids containing the SV40 origin of replication, thereby enhancing recombinant protein expression and viral production. HEK293T cells exhibit adherent epithelial morphology, robust growth rates, and high transfection efficiency, making them an ideal host for generating gene knockouts and for applications requiring transient or stable genetic manipulation.
DPYSL2 encodes collapsin response mediator protein 2 (CRMP2), a microtubule-associated protein that plays a central role in coordinating cytoskeletal dynamics. CRMP2 is regulated by phosphorylation through multiple upstream kinases, including glycogen synthase kinase-3?? (GSK-3??), cyclin-dependent kinase 5 (Cdk5), and Rho-associated coiled-coil kinase (ROCK), often downstream of guidance cues such as semaphorin-3A (Sema3A) and Reelin. Upon phosphorylation, CRMP2 dissociates from tubulin heterodimers and modulates microtubule polymerization, while simultaneously interacting with the Sra-1/WAVE1 complex and cofilin to influence actin filament reorganization. Additionally, CRMP2 binds kinesin-1 (KIF5) and Numb, linking it to intracellular trafficking and polarity determination. This multifunctional signaling node integrates input from pathways involving neuropilin-1, plexin-A, Fyn, and RhoA to control cellular responses like axon guidance, cell polarization, and migration.
Although HEK293T cells are non-neuronal, they provide a valuable platform for dissecting CRMP2-mediated cytoskeletal regulation and signaling independently of neuronal context. The DPYSL2 knockout in this cell background enables precise analysis of CRMP2??s role in fundamental processes such as cell adhesion, spreading, and motility, which are relevant to both developmental cell migration and cancer metastasis. Researchers can exploit the ease of transfection and protein expression in HEK293T cells to perform complementation studies, rescue experiments, and phospho-signaling analyses, thereby establishing structure?Cfunction relationships of CRMP2 domains and their interaction partners.
These polyclonal knockout cells are suitable for a range of experimental applications. In neurobiology-oriented cancer research, they can be used to model CRMP2-dependent invasive behavior, while in drug discovery they serve as a screening substrate for modulators of CRMP2 phosphorylation or interactions with tubulin and actin. Representative assays include Western blot analysis of total and phosphorylated CRMP2, RT-qPCR for residual DPYSL2 transcript, immunofluorescence staining of microtubule and actin networks, transwell migration and invasion assays, co-immunoprecipitation with tubulin or kinesin-1, and phospho-signaling profiling of GSK-3??, Cdk5, and Rho kinase. For additional technical details or to request this product, please contact Ascent Research.
