The DNAJC3 Knockout NCI-H1299 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that targets the DNAJC3 gene in the NCI-H1299 human lung adenocarcinoma cell line. This heterogeneous knockout pool provides a loss-of-function model to study DNAJC3 biology in a physiologically relevant cancer cell context.
Host cell background: NCI-H1299 is an epithelial cell line derived from a lymph node metastasis of a human non-small cell lung carcinoma (NSCLC). It is p53-deficient with wild-type KRAS and EGFR, representing a frequently used model for metastatic lung adenocarcinoma. The cell line exhibits robust proliferation and is amenable to genetic modifications, making it suitable for functional genomics studies.
DNAJC3 encodes an ER co-chaperone that negatively regulates the unfolded protein response (UPR) by inhibiting the kinases PERK (EIF2AK3) and PKR (EIF2AK2). Through direct interaction with PERK, it attenuates phosphorylation of eIF2??, reducing ATF4 translation and pro-apoptotic CHOP (DDIT3) induction. This activity integrates signals from upstream sensors IRE1, ATF6, and BiP/GRP78, and involves interactions with Hsp70 and IRE1, thereby fine-tuning the switch between adaptive survival and apoptosis under ER stress.
In the p53-deficient NCI-H1299 background, where compensatory survival signaling is essential for coping with oncogenic stress, DNAJC3-mediated PERK inhibition likely mitigates ER stress arising from hypoxia, nutrient deprivation, or protein misfolding. Disruption of DNAJC3 in this model enables investigation of UPR dysregulation effects on NSCLC cell survival, proliferation, migration, and sensitivity to chemotherapy, potentially uncovering novel therapeutic targets.
Typical research applications include monitoring UPR signaling dynamics via Western blot detection of BiP, phosphorylated PERK, phosphorylated eIF2??, and CHOP; quantifying XBP1 mRNA splicing and ATF4-dependent gene expression by RT-qPCR; and performing annexin V-based apoptosis assays alongside cell viability measurements under ER stress induction with tunicamycin or thapsigargin. The polyclonal knockout population also facilitates co-immunoprecipitation experiments to validate DNAJC3 interactions with PERK and other partners, and complementation assays to confirm phenotype specificity. For further technical information and ordering details, please contact Ascent Research.
