The DNAJC13 Knockout K-562 Polyclonal Cells constitute a CRISPR/Cas9-mediated polyclonal knockout cell population designed to disrupt the endogenous DNAJC13 gene in a K-562 human cell background. This product provides a mixed cell pool carrying heterogeneous gene-disruption events, enabling robust loss-of-function studies without clonal isolation. The knockout model supports investigation of endosomal protein sorting and retromer-dependent trafficking pathways. The polyclonal format reduces the risk of clonal artifacts and captures a broad range of genetic lesions, offering a versatile tool for biomedical research. By eliminating DNAJC13 expression, this model facilitates detailed interrogation of retrograde transport mechanisms and their impact on cellular homeostasis.
The host K-562 cell line is a well-established model derived from the pleural effusion of a chronic myelogenous leukemia (CML) patient in blast crisis. K-562 cells are BCR-ABL1-positive and exhibit the Philadelphia chromosome, making them a canonical platform for studying CML pathogenesis, erythroid differentiation, and apoptosis. Their lymphoblast-like morphology and suspension growth characteristics enable scalable experimental workflows. The K-562 background provides a human hematopoietic context in which to examine the crosstalk between oncogenic signaling and membrane trafficking processes. Consequently, DNAJC13 disruption in these cells is particularly relevant for dissecting the contributions of endocytic recycling to leukemic cell proliferation and survival.
DNAJC13 (RME-8) functions as a co-chaperone that cooperates with the retromer complex to sort integral membrane cargo receptors from early endosomes to the trans-Golgi network (TGN). Mechanistically, DNAJC13 directly interacts with the retromer subunits VPS35, VPS29, and VPS26, as well as sorting nexins SNX1 and SNX2, to drive retrograde transport. The protein also associates with HSP70 and DNAJB family co-chaperones, implicating ATP-dependent remodeling in cargo recognition. Key downstream cargoes controlled by DNAJC13 include Wntless (WLS) for Wnt morphogen secretion, the Notch receptor for regulated intramembrane proteolysis, and the cation-independent mannose-6-phosphate receptor (CI-MPR) for lysosomal enzyme delivery. Disruption of DNAJC13 therefore perturbs retrograde retrieval, leading to mislocalization of these cargoes, altered Wnt and Notch signaling output, and defects in endosome-to-TGN trafficking. Post-translational modifications, such as phosphorylation or ubiquitination, likely regulate DNAJC13 activity in response to endocytic demand, though specific upstream regulators remain to be defined.
In the K-562 leukemic environment, DNAJC13 knockout may uncover critical links between endosomal sorting and oncogenic phenotypes. Aberrant Wnt signaling is implicated in CML stem cell maintenance, and Notch pathway alterations can influence myeloid differentiation. By disrupting retromer function, this knockout model offers a means to probe how impaired WLS recycling affects Wnt ligand secretion and subsequent ??-catenin-dependent transcription in a BCR-ABL1-positive context. Additionally, defects in Notch receptor trafficking could modulate cell fate decisions and apoptosis sensitivity. Given the known association of DNAJC13 mutations with Parkinson??s disease, this cellular system also enables investigation of neurodegeneration-related trafficking defects in a non-neuronal host, facilitating comparative studies of retromer dysfunction across cell types. Thus, the DNAJC13 knockout K-562 cells serve as a specialized tool for examining the intersection of membrane trafficking, signal transduction, and malignant transformation.
This polyclonal knockout population is suitable for a wide range of experimental applications. Researchers can employ this model for endosomal trafficking studies using immunofluorescence and confocal microscopy to track the subcellular distribution of retromer components and cargo receptors. Western blotting and RT-qPCR enable confirmation of DNAJC13 disruption and assessment of downstream targets. Co-immunoprecipitation and interaction assays allow dissection of DNAJC13 complexes with SNX1/2, VPS35, and HSP70. Functional readouts include Wnt/Notch luciferase reporters, flow cytometry for surface receptor levels, and proliferation or apoptosis assays to evaluate biological impact. The cells are valuable for drug screening campaigns targeting trafficking defects or retromer-related pathologies. For technical inquiries, please contact Ascent Research.
