The DNAJC13 Knockout Raji Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Raji B lymphocyte line, designed to disrupt DNAJC13 gene function. This model provides a heterogeneous, loss-of-function system for studying DNAJC13 biology without relying on single-cell clones, allowing for the assessment of gene disruption effects across a genetically diverse pool of edited cells. The polyclonal format ensures robust experimental outcomes in endocytic trafficking and retromer function studies.
Raji cells are a human B lymphocyte line originating from a Burkitt’s lymphoma and are characterized by Epstein-Barr virus (EBV) positivity. These suspension cells are renowned for their capacity for high-level antibody production and are extensively used in immunological research to explore B cell activation, signaling, and immune responses. Their active endocytic and secretory pathways make them an ideal host for generating knockout models aimed at dissecting membrane trafficking and protein sorting mechanisms.
DNAJC13 encodes a co-chaperone that bridges protein folding and endosomal sorting. It interacts with HSP70 as part of the chaperone cycle and with the retromer complex subunits VPS35, SNX1, and SNX2, as well as the WASH complex component FAM21. Upstream regulators include the Rab5 and Rab7 GTPases and PI3P lipids. DNAJC13 facilitates retromer-dependent recycling of transmembrane proteins such as CI-MPR from endosomes to the trans-Golgi network, thereby preventing lysosomal degradation and promoting autophagic flux. This protein thus integrates quality control with cargo sorting.
In the Raji B lymphocyte context, DNAJC13 knockout is particularly relevant for examining retromer-mediated trafficking in cells with high biosynthetic and endocytic activity. Impairment of DNAJC13 is expected to disrupt CI-MPR retrieval, alter lysosomal function, and potentially affect immunoglobulin secretion and antigen presentation pathways. Moreover, given DNAJC13 associations with Parkinson??s disease and hereditary spastic paraplegia, this model offers a non-neuronal platform to investigate neurodegenerative disease mechanisms linked to endosomal dysfunction.
These polyclonal knockout cells support a wide array of experimental applications, including Western blotting for DNAJC13 and retromer components, RT-qPCR for mRNA knockdown verification, and flow cytometry to quantify surface CI-MPR recycling. Co-immunoprecipitation assays enable analysis of retromer complex integrity, while lysosomal activity and autophagic flux measurements provide functional readouts. Collectively, these assays facilitate research into endocytic trafficking, protein aggregation, and retromer biology. For additional information, contact Ascent Research.
