The DNM1 Knockout Raji Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population derived from Raji cells, targeting the DNM1 gene. This loss-of-function model enables investigation of dynamin-1??s role in clathrin-mediated endocytosis and membrane fission. The polyclonal nature ensures representation of diverse editing events, providing a robust system for studying gene disruption effects without clonal selection.
Raji cells are an immortalized B lymphocyte line originating from an Epstein-Barr virus (EBV)-positive Burkitt’s lymphoma patient. These cells are extensively employed in immunology and cancer research, serving as a model for B-cell malignancies and immune response studies. Their well-characterized growth properties and ease of manipulation make them a suitable host for gene-editing studies, particularly those investigating membrane trafficking pathways relevant to lymphocyte function.
Dynamin-1 (DNM1) is a large GTPase that oligomerizes at the necks of budding clathrin-coated vesicles. Upon GTP hydrolysis, it undergoes conformational changes that drive membrane fission, releasing synaptic vesicles. Its activity is regulated by phosphorylation and interactions with SH3 domain-containing proteins like endophilin and amphiphysin. Upstream regulators include calcineurin, GSK3??, the AP-2 complex, clathrin, and PIP2. Downstream, DNM1 facilitates vesicle uncoating via Hsc70 and auxilin, promoting emergence of early endosomes marked by Rab5. DNM1 also interacts with SNARE proteins and actin, linking endocytosis to cytoskeletal dynamics.
Although DNM1 is predominantly studied in neurons, its fundamental endocytic machinery is conserved, and its disruption in Raji cells provides a unique platform to dissect clathrin-mediated internalization in B lymphocytes. This knockout model is significant for studying how endocytic defects contribute to B-cell malignancies and neurodegenerative disorders like Alzheimer’s and Parkinson’s diseases. The Raji background allows exploration of DNM1-dependent signaling in an immune context, potentially revealing novel connections between membrane trafficking and oncogenic pathways.
Researchers can apply this polyclonal knockout cell population to assess endocytosis kinetics via transferrin uptake assays, visualize clathrin-coated pit formation by immunofluorescence, or quantify surface receptor internalization using flow cytometry. Biochemical analyses such as western blotting for DNM1 and GTPase activity assays enable validation of knockout efficiency and functional dissection. Phospho-signaling studies can elucidate regulatory mechanisms involving GSK3?? or calcineurin. This model is ideal for investigating synaptic vesicle recycling in a heterologous system and for screening modulators of endocytosis in cancer cells. For further technical details and ordering information, please contact Ascent Research.
