This product consists of a CRISPR/Cas9-edited polyclonal knockout cell population in which the DNAJC13 gene has been disrupted in A-549 cells. The polyclonal pool provides a heterogeneous population of gene-edited cells, enabling loss-of-function studies without requiring single-cell cloning. The CRISPR/Cas9-mediated gene disruption ablates DNAJC13 expression, eliminating the Hsp70 co-chaperone function critical for endosomal trafficking. This knockout model serves as a robust tool for investigating retromer-dependent cargo sorting and associated cellular pathways.
The host cell line A-549, derived from a 58-year-old Caucasian male lung adenocarcinoma, displays adherent epithelial morphology and alveolar type II pneumocyte-like characteristics, including surfactant protein expression. It harbors a KRAS G12S mutation and wild-type p53, with KRAS-driven signaling that makes it a relevant model for non-small cell lung cancer. Its genetic and phenotypic profile provides a suitable cellular context for investigating endosomal trafficking.
DNAJC13 functions as an Hsp70 co-chaperone on endosomes, critically facilitating retromer-dependent cargo retrieval. It interacts with retromer core subunits VPS35, VPS29, VPS26 and sorting nexins SNX1/SNX2, as well as RME-8 and FAM21, to sort cargo such as WLS, sortilin, and cathepsin D from early endosomes to the TGN. This process is essential for WNT signaling, autophagic flux, and lysosomal enzyme delivery. Stress-induced transcription factors ATF6 and XBP1 upregulate DNAJC13 under unfolded protein response conditions. Gene disruption therefore impedes endosome-to-Golgi transport, attenuating WNT target gene activation and autophagy.
In A-549 cells, DNAJC13 knockout permits dissection of retromer function and WNT signaling within the KRAS-mutant lung cancer environment. Although DNAJC13 mutations are linked to Parkinson??s disease and neuronal ceroid lipofuscinosis, its fundamental role in membrane traffic is cell-type independent. This model allows exploration of how retromer-mediated sorting impacts autophagic degradation and WNT output in a cancer-relevant background, potentially identifying context-dependent dependencies for therapeutic intervention.
The knockout pool supports applications in Parkinson??s disease modeling, endocytic trafficking, WNT/autophagy research, and drug screening. Typical assays include Western blotting for retromer components, immunofluorescence co-localization, co-immunoprecipitation, RT-qPCR of WNT targets, WNT reporter assays, transferrin uptake, and LC3 turnover measurements. The model is valuable for evaluating retromer stabilizers and lysosomal protease activity in neurodegenerative disease studies. For additional information or custom inquiries, please contact Ascent Research.
