The DNAJC14 Knockout HAP1 Polyclonal Cells are a heterogeneous population of HAP1 cells engineered via CRISPR/Cas9-mediated gene disruption to ablate DNAJC14 function. This polyclonal knockout format provides a loss-of-function model for studying DNAJC14-dependent processes without clonal selection, reflecting the genetic heterogeneity inherent to the edited population. The product is suitable for researchers requiring a robust knockout background for functional assays, pathway analysis, and pooled screening applications.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia cell line. It displays an adherent, fibroblast-like morphology and retains a largely haploid karyotype, making it a powerful model for genetic perturbation studies. The reduced genetic redundancy in HAP1 enables direct phenotypic assessment of gene disruptions, and its leukemic origin provides a relevant context for cancer biology and host?Cpathogen interaction research.
DNAJC14 encodes a J-domain co-chaperone that recruits HSPA8 (HSC70) to regulate its ATPase activity, a critical step for clathrin coat disassembly during clathrin-mediated endocytosis. DNAJC14 interacts with clathrin and auxilin (DNAJC6) to coordinate uncoating of clathrin-coated vesicles, and it is exploited by Rift Valley fever virus (RVFV) for replication, binding the viral nucleocapsid protein to facilitate ribonucleoprotein complex assembly. Upstream signals from the heat shock response and ER stress modulate DNAJC14 expression, positioning it at the nexus of protein quality control and membrane trafficking.
In the HAP1 near-haploid background, knockout of DNAJC14 disrupts HSPA8-dependent endocytosis and viral replication, providing a clean system to dissect these mechanisms. The model is particularly valuable for studying host factors essential for RVFV life cycle steps and for interrogating chaperone-driven processes in a leukemic context. Combined with the haploid genotype, this knockout allows for unambiguous assignment of phenotypes to DNAJC14 loss, facilitating screens for synthetic lethality or resistance mechanisms.
Key applications include investigating clathrin-mediated endocytosis via transferrin uptake assays, assessing viral replication kinetics through RVFV infection studies, and probing chaperone interactions using co-immunoprecipitation and immunofluorescence. The polyclonal population is also suited for pooled CRISPR screens and functional genomics experiments. Common readouts encompass western blotting, RT-qPCR, and flow cytometry for endocytic activity. For additional information or technical support, please contact Ascent Research.
