The DNAJC16 Knockout HAP1 Polyclonal Cells provide a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt DNAJC16 gene function in a near-haploid human cell model. This product comprises a genetically diverse pool of cells with targeted disruptions at the DNAJC16 locus, delivering a robust loss-of-function resource for probing protein homeostasis and ER stress biology. The polyclonal format captures editing heterogeneity, reducing clonal artifacts and enabling consistent population-level phenotypic readouts suitable for high-throughput screening and functional genomics assays.
HAP1 is a near-haploid adherent fibroblast-like cell line derived from KBM-7 chronic myeloid leukemia cells after spontaneous loss of chromosome 15. Its near-haploid karyotype permits efficient single-allele gene disruption, making it a favored platform for CRISPR-based knockout studies. HAP1 cells are widely used in functional genomics, genetic screens, and drug sensitivity profiling due to their well-characterized proteome and transcriptome. The hematopoietic lineage background provides relevance to leukemia research and other cancer types, while the haploid state simplifies the interpretation of loss-of-function phenotypes.
DNAJC16 encodes an ER-resident DnaJ/Hsp40 co-chaperone that stimulates the ATPase activity of HSP70 chaperones, most notably HSPA5/BiP. Integral to the heat shock protein cycle, DNAJC16 drives protein folding, translocation, and ER-associated degradation (ERAD) of misfolded clients. Its expression is controlled by key ER stress transcription factors ATF6, XBP1, and HSF1. The co-chaperone interacts directly with HSP70 family members, nucleotide exchange factors, and unfolded polypeptides, influencing downstream UPR effectors such as CHOP and GADD34. Thus, DNAJC16 links chaperone-mediated quality control to adaptive and apoptotic ER stress signaling.
Knockout of DNAJC16 in HAP1 cells disrupts HSP70-driven protein quality control, causing accumulation of misfolded proteins and heightened ER stress sensitivity. The haploid advantage amplifies the phenotypic impact of single-gene loss, facilitating clear dissection of UPR pathway activation and ERAD efficiency. Upon challenge with ER stressors such as tunicamycin or thapsigargin, these cells may exhibit hyperactivation of the PERK-eIF2??-ATF4 and IRE1-XBP1 branches. Consequently, this model is instrumental for studying DNAJC16 in cancer cell adaptation, neurodegeneration-linked protein aggregation, and responses to proteostasis-targeting drugs.
This knockout population supports diverse cell-based assays. Standard applications include western blotting for UPR markers (BiP, CHOP), RT-qPCR of ER stress target genes, and viability assays under ER stress conditions. Flow cytometry with annexin V quantifies apoptosis, while co-immunoprecipitation of HSP70 reveals chaperone-client interaction changes. These cells are also amenable to high-content screening and reporter assays for ER stress response elements, enabling functional genomics screening and pharmacological profiling of modulators targeting the proteostasis network. For additional technical details, please contact Ascent Research.
