The DNAJB11 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population designed for ER stress and functional genomics research, consisting of a heterogeneous pool of HAP1 cells with disrupted DNAJB11. This polyclonal format provides a robust model for studying DNAJB11-dependent processes without clonal artifacts, suitable for high-throughput screening and pathway analysis in a near-haploid genetic background that facilitates unambiguous genotype-phenotype correlations.
The HAP1 cell line is a near-haploid human cell line derived from KBM-7 chronic myeloid leukemia, widely used for functional genomics and knockout studies due to its stable haploid karyotype and genetic tractability. Its simplified genetic landscape enhances penetrance of gene disruptions, making it ideal for studying dosage-sensitive pathways such as the unfolded protein response (UPR) and for large-scale CRISPR screens.
DNAJB11 encodes an ER co-chaperone with a J-domain that recruits HSPA5/BiP to misfolded proteins for folding and ER-associated degradation (ERAD). It interacts with OS9, SEL1L, and the HRD1/SYVN1 ligase complex, and its expression is regulated by ER stress sensors ATF6, IRE1/XBP1, and PERK/ATF4. DNAJB11 also modulates UPR by attenuating IRE1/XBP1 and associates with HSP90B1/GRP94 and VCP/p97. Knockout disrupts ER quality control, causing misfolded protein accumulation and compensatory UPR activation, including PERK-mediated eIF2?? phosphorylation and upregulation of ATF4 and CHOP.
In HAP1 cells, DNAJB11 ablation chronically activates ER stress sensors and downstream effectors, providing a tractable system to dissect UPR signaling dynamics and ERAD substrate flux. This model recapitulates molecular features of ADTKD and broader ER stress-related pathologies in cancer and neurodegeneration. The haploid background enables detection of subtle UPR phenotypes and dose-response studies with tunicamycin or ER stress inhibitors, while the polyclonal composition reduces background-specific bias, offering a reliable platform for benchmarking ER proteostasis assays and evaluating chemical chaperones.
Applications include UPR dissection via Western blotting for p-eIF2??, CHOP, and XBP1s; RT-qPCR for ER stress targets; immunofluorescence for ER morphology; flow cytometry for apoptosis (Annexin V/PI); co-immunoprecipitation of HSPA5; tunicamycin sensitivity; and proteasome activity assays. These cells support functional genomics screens, drug target validation for protein misfolding diseases, and chaperone biology research. Contact Ascent Research for further information.
