The DNM1 Knockout HeLa Polyclonal Cells comprise a CRISPR/Cas9-edited polyclonal knockout population from the HeLa cervical adenocarcinoma line, wherein DNM1 disruption abrogates dynamin-1 protein expression. This loss-of-function system facilitates investigation of clathrin-dependent endocytosis without remnant dynamin activity. The polyclonal composition yields a genetically diverse pool, ideal for pooled screening and bulk endocytic assays.
HeLa cells, the first immortalized human cell line derived from cervical adenocarcinoma, serve as a cornerstone model in cell biology. These epithelial cells maintain cervical barrier functions and active endomembrane trafficking. Their rapid growth and amenability to genetic engineering have established HeLa as a prime system for studies of receptor signaling, membrane dynamics, and oncogenesis. In this context, DNM1 knockout permits precise functional dissection of dynamin-1 in a cancerous epithelial setting.
Dynamin-1 (DNM1) encodes a mechanochemical GTPase that drives membrane scission during clathrin-mediated endocytosis, a process fundamental to synaptic vesicle recycling and receptor internalization. The protein is recruited to clathrin-coated pits by binding partners amphiphysin, endophilin, and syndapin, oligomerizing into helical rings. GTP hydrolysis triggers conformational changes that constrict and sever the membrane neck. Its activity is dynamically regulated by Ca2+ influx and phosphorylation/dephosphorylation by Cdk5 and calcineurin, respectively. Downstream, dynamin-1 mediates internalization of transferrin receptor (TfR) and AMPA-type glutamate receptors (AMPAR), thereby modulating surface receptor abundance. Additionally, it collaborates with the AP-2 adaptor complex and clathrin heavy chain to select cargo and sculpt vesicles, positioning it as a central coordinator of receptor-mediated endocytosis.
In the HeLa adenocarcinoma model, dynamin-1 disruption offers a clinically pertinent system to study how endocytic alterations affect cancer cell signaling, receptor desensitization, and drug responses. The low endogenous dynamin-1 expression in HeLa cells permits unambiguous attribution of endocytic defects to the knockout. This polyclonal knockout pool is particularly suited for investigating heterogeneity in clathrin-coated pit dynamics and compensation by dynamin-2.
Researchers can leverage this knockout model in a wide range of assays, including transferrin uptake measurements to assess bulk clathrin-mediated endocytosis, immunofluorescence microscopy to examine clathrin-coated pit morphology and dynamin-interacting protein localization, and live-cell imaging with pH-sensitive probes such as pHrodo to track vesicle acidification and trafficking dynamics. Western blotting for dynamin-1 confirms knockout efficiency, while flow cytometry-based receptor internalization assays provide high-throughput analysis of endocytic uptake. The polyclonal nature facilitates investigation of nanoparticle uptake heterogeneity for drug delivery refinement and comparative studies of viral entry mechanisms. For further information or to request a quote, please contact Ascent Research.
