The DNAJC3 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal population in which the DNAJC3 gene has been disrupted, generating a loss-of-function model for investigating the regulatory roles of the P58IPK chaperone. This product provides a heterogeneous pool of edited cells, ideal for bulk functional studies of DNAJC3-dependent cellular processes. The polyclonal format captures the spectrum of CRISPR-induced mutations across the cell population, enabling robust assessment of biological phenotypes without clonal selection bias.
The parental HEK293T cell line is a widely utilized human embryonic kidney cell derivative, stably expressing the SV40 large T antigen. This modification promotes episomal replication of plasmids containing the SV40 origin, facilitating high-level transient and stable protein expression. HEK293T cells are a standard model for mechanistic cell biology, signal transduction research, and protein interaction studies, making them a versatile host for gene knockout applications.
DNAJC3 (P58IPK) is an ER stress-inducible negative regulator of the PERK-eIF2??-ATF4 branch of the unfolded protein response (UPR). Upstream stimuli such as tunicamycin, thapsigargin, hypoxia, or glucose deprivation induce its expression through ATF4, XBP1, and CHOP. The protein directly binds and inhibits PERK (EIF2AK3) kinase activity, decreasing eIF2?? phosphorylation and subsequent ATF4 translation. This reduces transcription of pro-apoptotic CHOP and GADD34, while its interactions with Hsp70 and BiP/GRP78 further coordinate chaperone function. The DNAJC3-PERK feedback loop is critical for promoting cell survival under moderate ER stress.
In the HEK293T background, disruption of DNAJC3 is predicted to relieve the inhibitory tone on PERK, resulting in enhanced eIF2?? phosphorylation even under basal conditions and hyperactivation of the PERK arm upon ER stress. This knockout model enables precise dissection of PERK-dependent signaling without pharmacological modulation, facilitating studies of how sustained UPR activation affects cell fate, protein synthesis, and stress adaptation. The polyclonal nature of the population ensures that observed phenotypes reflect the average behavior of multiple genotypes, reducing the risk of clonal artifacts.
These cells are ideal for Western blotting of PERK pathway markers (phospho-eIF2??, ATF4, CHOP), RT-qPCR for UPR target genes, co-immunoprecipitation of DNAJC3-PERK complexes, and cell viability assays under ER stress inducers such as tunicamycin. They support drug screening for UPR modulators and disease modeling for ER stress-related pathologies. For further details, please contact Ascent Research.
