The DNAJC3 Knockout K-562 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the K-562 human leukemia cell line. This heterogeneous pool carries disruptions in the DNAJC3 gene, enabling bulk loss-of-function studies without clonal selection artifacts. The product offers a robust model to interrogate DNAJC3 function in endoplasmic reticulum stress and unfolded protein response (UPR) signaling.
K-562 is an immortalized cell line established from a female chronic myelogenous leukemia patient in blast crisis. Philadelphia chromosome positive, it expresses BCR-ABL fusion kinase and serves as a standard model for CML biology, hematopoietic differentiation, and drug resistance. The high protein secretion demand of this lymphoblastoid line renders it sensitive to ER stress, making it an appropriate host for UPR gene knockout investigations.
DNAJC3 encodes p58IPK, an ER co-chaperone that regulates BiP/GRP78 (HSPA5) ATPase activity and inhibits PKR (EIF2AK2) to control eIF2?? phosphorylation and translational attenuation. Transcriptionally induced by ATF4, XBP1, and ATF6 under stress, DNAJC3 modulates PERK and IRE1 pathways and interacts with BiP, PKR, PERK, IRE1, and HSPA8. Knockout disrupts UPR signaling, altering eIF2??, ATF4, CHOP expression, and apoptosis susceptibility.
In the K-562 leukemia background, DNAJC3 loss may exacerbate ER stress sensitivity, potentially affecting survival signals and drug responses. This model enables examination of UPR?CBCR-ABL cross-talk, apoptosis regulation, and translational control mechanisms. Researchers can assess how DNAJC3 deficiency influences phosphorylation of eIF2?? and activation of downstream stress effectors in a relevant oncogenic context.
Typical assays include Western blotting for BiP, phospho-eIF2??, ATF4, CHOP; RT-qPCR for XBP1 splicing; flow cytometry for apoptosis (Annexin V/PI); and cell viability assays (MTT) under tunicamycin or thapsigargin treatment. Co-immunoprecipitation and RNA-seq can further resolve altered protein networks and transcriptomes. For technical inquiries, contact Ascent Research.
