DNAJB2 Knockout HeLa Polyclonal Cells are a human cervical adenocarcinoma knockout cell product generated by CRISPR/Cas9-mediated disruption of the DNAJB2 gene in the HeLa cell line. This product is supplied as a polyclonal knockout cell population, meaning it comprises a heterogeneous mixture of HeLa cells carrying various CRISPR-induced alleles at the DNAJB2 locus, providing a loss-of-function model without clonal isolation. The pooled population retains the inherent genetic diversity of the editing process and is suitable for studying bulk protein quality control effects.
HeLa cells are an immortalized human cervical cancer cell line positive for HPV18, derived from a cervical adenocarcinoma. Widely used in research, these cells provide a robust epithelial model for studying proteostasis and chaperone functions.
DNAJB2 encodes a co-chaperone regulating Hsp70 ATPase activity. It directly interacts with HSPA8/Hsc70 and HSPA1A/Hsp70 to target misfolded or aggregation-prone client proteins, such as SOD1 and Tau. DNAJB2 partners with the E3 ubiquitin ligase STUB1/CHIP and BAG family co-chaperones to facilitate substrate ubiquitination and proteasomal degradation. Additionally, it participates in autophagy-mediated clearance of protein aggregates via associations with p62/SQSTM1. Upstream regulators include heat shock factor 1 (HSF1) and the unfolded protein response sensors IRE1, PERK, and ATF6, integrating DNAJB2 into cellular stress responses.
In HeLa cells, DNAJB2 knockout compromises the protein quality control network, leading to accumulation of polyubiquitinated proteins and formation of insoluble aggregates, which can be detected by filter trap assays and immunofluorescence. The loss of DNAJB2 impairs both proteasomal and autophagic degradation routes, simulating proteotoxic stress conditions relevant to neurodegenerative disorders like Charcot-Marie-Tooth disease type 2T and spinal muscular atrophy. This model allows dissection of DNAJB2-specific functions in HSP70 cycle regulation and degradation pathway selection, providing a platform to explore the interplay between chaperone-mediated folding, ER-associated degradation, and autophagy in a cancer cell background.
Typical research applications include mechanistic studies of protein aggregation diseases, screening of small molecules that modulate proteostasis, and investigation of co-chaperone-dependent client processing. Researchers can employ western blotting to assess DNAJB2, HSP70, and ubiquitinated protein levels; RT-qPCR to monitor transcriptional responses; proteasome activity and autophagy flux assays to evaluate degradation pathways; and cellular thermal shift assays (CETSA) to probe chaperone interactions. This polyclonal knockout population is a versatile tool for functional genomics and drug discovery targeting proteostasis networks. For additional information or custom inquiries, please contact Ascent Research.
