The DNAJC7 Knockout NCI-H1299 Polyclonal Cells product provides a CRISPR/Cas9-edited polyclonal knockout cell population in which the DNAJC7 gene has been disrupted in NCI-H1299 cells. This polyclonal pool serves as a heterogeneous loss-of-function model for investigating DNAJC7-dependent proteostasis and stress response mechanisms.
The host NCI-H1299 cell line is a human lung adenocarcinoma line derived from lymph node metastasis. It is TP53 null and grows as an adherent epithelial monolayer, making it a widely used model for p53-independent cancer biology and stress response studies.
DNAJC7 encodes a J-domain co-chaperone that stimulates the ATPase activity of heat shock protein 70 (HSP70) isoforms, primarily HSPA1A and HSPA8. DNAJC7 transcription is induced by heat shock factor 1 (HSF1) in response to proteotoxic stress, and the protein acts by recruiting HSP70 to client polypeptides, where it collaborates with BAG family co-chaperones and the E3 ubiquitin ligase STUB1/CHIP to determine client fate. BAG proteins facilitate nucleotide exchange, while STUB1/CHIP ubiquitinates terminally misfolded clients for proteasomal degradation. Disruption of DNAJC7 disrupts this coordination, impairing HSP70 cycling and leading to accumulation of unfolded client proteins and cytotoxic aggregates. Consequently, cellular proteostasis is compromised, and stress signaling pathways are dysregulated.
In the NCI-H1299 lung adenocarcinoma background, which is TP53 null and therefore deficient in p53-mediated stress responses, DNAJC7 knockout may further sensitize cells to proteotoxic insults, revealing critical dependencies on chaperone-mediated quality control. This model permits investigation of synthetic lethal interactions with proteasome inhibitors (e.g., bortezomib) or HSP90 inhibitors, and the adherent epithelial morphology facilitates high-content imaging of protein aggregation and stress granule dynamics.
Typical applications include investigating chaperone function and proteostasis, western blotting for HSP70 and client proteins, proteasome activity assays, cell viability assays under heat shock or proteasome inhibition, immunofluorescence detection of aggregates, and co-immunoprecipitation of chaperone complexes. The polyclonal knockout population is also suitable for functional genomics screens and drug sensitivity profiling in lung cancer research. For further information, please contact Ascent Research.
