The DNM1 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population in which the DNM1 gene has been disrupted to impair expression of Dynamin 1. This population provides a genetically heterogeneous loss-of-function model, enabling robust assessment of Dynamin 1-dependent processes without clonal selection artifacts. The polyclonal format facilitates the study of gene disruption effects across a range of editing outcomes, offering a versatile tool for functional genomics and endocytosis research in a widely used human cell background.
These cells are derived from the HEK293T host cell line, a female embryonic kidney epithelial derivative of HEK293 that stably expresses the SV40 large T-antigen. This immortalized line is renowned for its high transfectability, robust protein expression, and capacity for efficient viral packaging, making it a preferred system for studying membrane trafficking and signaling. The adherent epithelial morphology and rapid growth kinetics of HEK293T cells support high-throughput and imaging-based assays, ensuring compatibility with standard cell culture workflows.
Dynamin 1, encoded by DNM1, is a large GTPase essential for membrane fission during clathrin-mediated endocytosis and synaptic vesicle recycling. It assembles into helical polymers at the necks of budding clathrin-coated vesicles, where GTP hydrolysis drives constriction and scission. Dynamin 1 function is regulated by interactions with amphiphysin, endophilin, and syndapin, and is modulated by AP-2 complex, calcium signaling, and Src-family kinases. Downstream, Dynamin 1 acts on clathrin-coated vesicles to internalize cargo such as transferrin receptor and epidermal growth factor receptor (EGFR), directly linking receptor-mediated uptake to intracellular signaling cascades. Additional interacting partners like Grb2, cortactin, and SNX9 fine-tune Dynamin 1 localization and activity at distinct endocytic sites.
Although DNM1 is predominantly associated with neuronal functions, its fundamental role in clathrin-mediated endocytosis is conserved across cell types. In HEK293T cells, Dynamin 1 contributes to constitutive and growth factor?Cinduced endocytosis, and its knockout reveals functional compensation by the ubiquitously expressed dynamin isoforms DNM2 and DNM3. This model is therefore valuable for dissecting isoform-specific roles in membrane trafficking and for identifying pathway components that become rate-limiting upon loss of Dynamin 1. The knockout cells offer a genetically defined platform to explore how endocytic defects impact signaling downstream of EGFR and transferrin receptor, with relevance to neurodevelopmental disorders linked to DNM1 mutations.
Researchers can employ this polyclonal knockout model in an array of functional assays, including transferrin uptake assays to quantify clathrin-mediated endocytosis, Western blotting and immunofluorescence to assess Dynamin 1 depletion and vesicle marker distribution, and live-cell imaging to monitor vesicle dynamics. The cells are suitable for GTPase activity measurements and electron microscopy studies of clathrin-coated pit morphology, as well as for drug discovery screens targeting dynamin-related pathways. This product thus supports mechanistic investigations into membrane trafficking, receptor signaling, and the cellular consequences of DNM1 loss. For further information or to discuss your specific research needs, please contact Ascent Research.
