The DNAJC15 Knockout HAP1 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed for functional investigation of DNAJC15 loss in a human near-haploid myeloid leukemia background. This polyclonal pool derives from the HAP1 cell line and provides a genetically disrupted DNAJC15 model suitable for studying mitochondrial regulation, metabolic reprogramming, and disease-relevant pathways without clonal selection bias.
HAP1 is an adherent, near-haploid human cell line originally derived from the chronic myeloid leukemia line KBM-7. Its near-haploid karyotype simplifies the interpretation of gene knockout phenotypes by reducing genetic redundancy, making it an ideal host for systematic functional genomics. Widely adopted in haploid genetic screens and leukemia research, HAP1 cells maintain rapid proliferation and robust metabolic activity, providing a consistent background for studying mitochondrial and cancer-related processes.
DNAJC15 encodes a mitochondrial co-chaperone that inhibits respiratory chain complex I and modulates the Hsp70 chaperone cycle. Mechanistically, DNAJC15 interacts directly with HSPA9 (mtHsp70) and TIMM23 translocase components to regulate mitochondrial protein import, while its association with NDUFS1 and MT-ND1 suppresses complex I activity. This inhibition decreases oxidative phosphorylation and promotes a glycolytic metabolic state. DNAJC15 expression is silenced by DNA methylation in certain cancers and is regulated by estrogen signaling, PGC-1??, and metabolic stress, linking its function to metabolic adaptation and tumorigenesis. Downstream consequences of its loss include enhanced complex I activity, elevated ATP synthesis via ATP5A1, and altered glycolytic enzyme expression.
In the HAP1 cellular context, DNAJC15 knockout is predicted to relieve complex I inhibition, leading to increased respiration and potentially altered chemosensitivity. The near-haploid background facilitates clear genotype?Cphenotype correlations in assays measuring mitochondrial function and drug response. This model is particularly suited for investigating the role of DNAJC15 in breast and ovarian cancer metabolic reprogramming, obesity, and metabolic syndrome, as well as for exploring synthetic lethal interactions via haploid genetic screens.
Researchers can employ these polyclonal knockout cells in a range of applications, including immunoblotting for DNAJC15 and complex I subunits, Seahorse respirometry to assess oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), ATP luminescence assays, complex I enzymatic activity measurements, and cisplatin sensitivity profiling. RT-qPCR can confirm transcript loss, while interaction studies with HSPA9 and NDUFS1 can be conducted. These cells enable dissection of mitochondrial function, cancer metabolism, and chemoresistance mechanisms. For additional details, please contact Ascent Research.
