DYNAP Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population targeting the DYNAP gene in the HAP1 near-haploid human cell line. This heterogeneous pool of cells carries gene disruptions introduced by CRISPR/Cas9, enabling loss-of-function studies without isolating clonal lines. The polyclonal format permits population-level analysis of DYNAP-dependent phenotypes, suitable for high-throughput screening and pathway studies.
HAP1 cells derive from KBM-7, a near-haploid chronic myeloid leukemia line, offering a simplified genetic background with only one allele per gene, which facilitates efficient CRISPR/Cas9-mediated knockout generation. Widely used for functional genomics and haploid genetic screens, HAP1 cells exhibit stable growth and are compatible with arrayed and pooled screening formats, making them ideal for systematic gene perturbation studies.
DYNAP encodes an endocytic accessory protein that binds dynamin-1 and stimulates its GTPase activity, promoting oligomerization required for membrane scission in clathrin-mediated endocytosis. DYNAP activity is regulated by upstream signals including EGF, synaptic activity, and calcium signaling, and it interacts with clathrin heavy chain (CLTC), the AP-2 complex (AP2M1), and amphiphysin (AMPH) to coordinate dynamin-1 (DNM1) recruitment at endocytic sites. Through these interactions, DYNAP facilitates efficient synaptic vesicle recycling and membrane trafficking. Loss of DYNAP disrupts dynamin-1 activation, impairing endocytic vesicle formation.
The HAP1 knockout model provides a clear system to dissect DYNAP??s role in clathrin-mediated endocytosis and synaptic vesicle cycling due to minimal genetic redundancy. This model is valuable for investigating neurological disorders such as epilepsy and neurodevelopmental disorders, where defective endocytosis contributes to pathology. Researchers can monitor how DYNAP loss affects dynamin-1-dependent membrane fission and downstream trafficking events.
Applications include studying endocytic trafficking mechanisms, screening for regulators of clathrin-mediated endocytosis, and modeling synaptic dysfunction. Compatible assays include western blotting, immunofluorescence, transferrin uptake, FM dye release, and co-immunoprecipitation to validate knockout efficiency and probe protein interactions. These cells support functional interrogation of DYNAP in endocytosis and offer a platform for drug discovery targeting membrane trafficking. For further information and technical support, contact Ascent Research.
