DNAJC27 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population that disrupts the DNAJC27 gene in the near-haploid human HAP1 cell line. This product offers a heterogeneous pool of loss-of-function alleles, providing a cost-effective model for functional genomics and high-throughput screening without the need for clonal isolation. It serves as a powerful tool to study DNAJC27-dependent processes such as protein quality control, endosomal trafficking, and chaperone-mediated autophagy.
HAP1 cells are a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia (CML) blast crisis isolate. Their largely haploid karyotype enables efficient gene disruption and straightforward genotype-phenotype correlations, making them a standard model for genetic screens. Retaining key cancer signaling pathways and protein homeostasis machinery, HAP1 is ideal for dissecting chaperone networks and stress responses in a cancer-relevant context.
DNAJC27 encodes a DnaJ domain-containing co-chaperone that stimulates Hsp70 ATPase activity, promoting substrate folding or degradation. Its N-terminal Rab-like domain suggests additional roles in membrane tethering for vesicular trafficking. DNAJC27 interacts with HSPA1A, HSPA8, BAG3, and STUB1/CHIP, linking the Hsp70 cycle to autophagy and endosomal sorting. Upstream, HSF1 and ER stress sensors induce DNAJC27 under proteotoxic and nutrient stress; downstream, it modulates Hsp70 substrate processing, autophagic cargo recognition via LAMP2A, and endosomal Rab GTPase activity.
In HAP1 cells, DNAJC27 knockout amplifies phenotypes due to the near-haploid genome, facilitating detection of subtle effects on protein aggregation, apoptosis, and drug sensitivity. This model is particularly suited to study how DNAJC27 integrates chaperone-mediated autophagy, endosomal trafficking, and cancer cell fitness, with relevance to asthenozoospermia and cancers where chaperone networks are dysregulated. The Rab-like domain??s role in membrane dynamics can also be probed in ciliogenesis and vesicular transport contexts.
Typical applications include western blotting, RT-qPCR, immunofluorescence, and flow cytometry to monitor protein aggregation and DNAJC27 interactions via co-immunoprecipitation. HSP70 ATPase assays, cell viability, and apoptosis profiling are readily performed. The polyclonal population excels in high-throughput screens for modulators of protein quality control, drug sensitivity profiling, and ciliogenesis studies. For technical support, please contact Ascent Research.
