The DNM3 Knockout HeLa Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the human DNM3 gene in the HeLa epithelial cell line. This product provides a heterogeneous loss-of-function model for studying dynamin 3-dependent cellular processes. Generated through CRISPR/Cas9-mediated gene disruption, the polyclonal pool encompasses a range of editing outcomes, offering a robust tool for endocytosis and cytoskeletal dynamics research without the biases associated with clonal selection. As a polyclonal knockout product, it is well-suited for experiments requiring a mixed population to capture phenotypic variability in membrane trafficking and migration assays.
HeLa cells, derived from a cervical adenocarcinoma and positive for HPV18, are an immortalized human epithelial line widely employed in cancer biology, cell signaling, and endocytosis studies. Their highly proliferative nature, robust adhesion, and well-documented migration behavior make them an ideal background for investigating the roles of dynamin 3 in clathrin-mediated endocytosis and actin remodeling. The extensive characterization of HeLa cells ensures compatibility with existing literature and facilitates direct comparisons across gene perturbation experiments targeting endocytic machinery.
Dynamin 3 is a large GTPase that orchestrates membrane fission during clathrin-mediated endocytosis and integrates actin filament dynamics through its interactions with multiple partners. It forms complexes with amphiphysin, endophilin, and intersectin, and is regulated by upstream kinases such as Src and PKC, as well as the small GTPases Rac1 and Cdc42. DNM3 also binds Grb2 and profilin, linking membrane scission to actin polymerization. Its GTPase activity is essential for the internalization of cargo like transferrin receptor, and in non-neuronal cells, it drives receptor trafficking and focal adhesion turnover, influencing cell motility.
Disruption of DNM3 in HeLa cells impairs dynamin-dependent membrane fission, leading to defective clathrin-mediated endocytosis and altered trafficking of receptors such as the transferrin receptor. This knockout model exhibits attenuated filopodia formation and reduced cell migration, consistent with DNM3??s role in actin reorganization at the leading edge. The polyclonal nature of this product may reveal variable phenotypic penetrance, making it a valuable system for probing the interplay between endocytosis and cytoskeletal remodeling in cancer cell motility. It also enables studies on how heterogeneous loss of DNM3 affects metastatic potential and focal adhesion dynamics.
These polyclonal knockout cells are ideally suited for a range of functional assays, including transferrin uptake measurements, immunofluorescence microscopy with phalloidin staining, transwell migration assays, and co-immunoprecipitation of interacting proteins such as amphiphysin and endophilin. Additional applications encompass drug uptake and nanoparticle internalization studies, RNA-seq to profile transcriptional responses, and GTPase activity assays. The model supports investigations into endocytic pathway regulation and the role of dynamin 3 in cancer metastasis. For further information or to discuss specific experimental requirements, please contact Ascent Research.
