The DNAJC13 Knockout HEK293T Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population targeting DNAJC13 in the HEK293T background. This cell pool enables loss-of-function studies of DNAJC13, a co-chaperone involved in retromer-mediated endosomal trafficking. Through CRISPR/Cas9-mediated gene disruption, the population is suitable for investigating the functional consequences of DNAJC13 ablation without the need for clonal isolation.
HEK293T cells are a widely employed human embryonic kidney epithelial cell line stably expressing the SV40 large T-antigen, which permits episomal replication of plasmids containing the SV40 origin. Known for high transfectability, they are a standard platform for recombinant protein expression, lentiviral and retroviral packaging, and mechanistic cell biology studies.
DNAJC13 encodes a J-domain co-chaperone that partners with Hsc70 to regulate endosomal membrane dynamics and cargo sorting. It functions within the retromer-mediated retrieval pathway, interacting directly with retromer components VPS35, VPS26, and VPS29, as well as sorting nexins SNX1 and SNX2, and Rab GTPases Rab7 and Rab9. This complex facilitates the retrograde trafficking of transmembrane proteins such as the cation-independent mannose 6-phosphate receptor (CI-MPR) and Wntless (WLS) from endosomes to the trans-Golgi network. Upstream regulators include phosphatidylinositol 3-phosphate and Rab7/Rab9, while downstream disruption of DNAJC13 impairs CI-MPR recycling and WLS transport, attenuating Wnt signaling and causing endosomal morphological defects.
In the HEK293T background, which retains functional endosomal and retromer pathways, DNAJC13 knockout provides a physiologically relevant model to dissect retromer-dependent sorting. Given that autosomal dominant mutations in DNAJC13 are associated with Parkinson disease 21 (PARK21), this polyclonal knockout population serves as a robust tool for investigating pathogenic mechanisms underlying Parkinson disease, including aberrant Wnt signaling and cargo mis-sorting.
This knockout cell product is suited for a range of experimental applications, including characterization of retromer-mediated trafficking using co-immunoprecipitation of DNAJC13 with Hsc70 and retromer subunits, quantitative recycling assays for CI-MPR, and immunofluorescence microscopy to visualize endosomal morphology changes. DNAJC13 disruption can be combined with Wnt pathway reporters (TOP/FOP) to assess signaling output, and the polyclonal nature supports bulk analysis by Western blotting and RT-qPCR. The model also enables high-throughput screening for small molecules that restore retromer function. For additional information or technical support, please contact Ascent Research.
