The DNAJC10 Knockout A-549 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout population derived from the A-549 human lung adenocarcinoma cell line. This product serves as a versatile tool for dissecting the roles of DNAJC10, also known as ERdj5, an ER-resident co-chaperone and disulfide reductase critical for ER-associated degradation (ERAD) of misfolded glycoproteins. The polyclonal nature of the knockout ensures a diverse genetic background suitable for population-level studies of ER stress and proteostasis.
The host A-549 cell line, originally established from a 58-year-old male lung adenocarcinoma, bears an activating KRASG12S mutation and exhibits alveolar type II epithelial characteristics. Widely used as a model for human lung adenocarcinoma, A-549 cells provide a biologically relevant context for investigating tumor cell dependence on protein quality control pathways, particularly in the setting of KRAS-driven malignancies.
DNAJC10/ERdj5 functions within the ER lumen by reducing disulfide bonds in misfolded glycoproteins, a prerequisite for substrate recognition by EDEM1 and subsequent handoff to the SEL1L?CHRD1 retrotranslocation complex, which includes Derlin-1 and OS9. Its expression is transcriptionally upregulated by ER stress sensors such as IRE1-mediated XBP1s and ATF6, linking it directly to the unfolded protein response (UPR). Key intermolecular partners encompass BiP/GRP78, VIMP, and p97/VCP, and its activity facilitates the dislocation of ERAD substrates like mutant ??1-antitrypsin Z and viral NS1. Disruption of DNAJC10 therefore stalls disulfide processing, blocking efficient degradation.
In the context of KRAS-mutant A-549 cells, DNAJC10 knockout cripples the ERAD machinery, provoking accumulation of misfolded proteins and sustained ER stress. This can alter cellular outcomes such as proliferation, apoptosis, and sensitivity to chemotherapeutics. The model is pertinent for exploring how lung adenocarcinoma cells manage proteotoxic insults and may reveal exploitable therapeutic vulnerabilities. Given the broader involvement of ERAD and UPR in neurodegeneration, viral infection, and metabolic disorders, the cells extend utility beyond tumor biology.
These polyclonal knockout cells support diverse applications including monitoring UPR activation via western blotting for GRP78, CHOP, and ATF4; measuring ERAD substrate turnover by cycloheximide chase; and assessing cell viability under ER stressors like tunicamycin or thapsigargin. They are further suited for viral replication studies, proteasome inhibitor sensitivity profiling, immunofluorescence localization of ERAD complexes, co-immunoprecipitation of interactors, and migration/invasion assays. For additional information, please contact Ascent Research.
