DNAJA2 Knockout HEK293T Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the widely used HEK293T human embryonic kidney cell line. This product features targeted disruption of the DNAJA2 gene, generating a loss-of-function model for investigating the role of the DNAJA2 co-chaperone in protein quality control and cellular stress responses. The polyclonal format preserves the natural genetic diversity of the edited cell pool, enabling researchers to analyze population-level phenotypes without clonal selection biases.
The host cell line, HEK293T, is a robust and versatile system extensively employed in biomedical research. These cells constitutively express the SV40 large T antigen, which promotes episomal replication of plasmids containing the SV40 origin of replication, leading to high transgene expression and efficient viral vector production. Their human origin and well-characterized molecular pathways make them particularly suitable for studying protein homeostasis, signal transduction, and stress biology.
DNAJA2 is a member of the DnaJ/Hsp40 family of co-chaperones that plays a critical role in coordinating Hsp70 chaperone activity. Mechanistically, DNAJA2 binds unfolded or misfolded protein substrates and delivers them to Hsp70 for ATP-dependent refolding, or to the ubiquitin-proteasome system for degradation. Upstream, DNAJA2 expression is induced by heat shock transcription factor HSF1 in response to cellular stresses such as heat shock and oxidative stress. DNAJA2 functionally interacts with Hsp70, the proteasome, and other DnaJ family members, positioning it at the intersection of protein folding, ER-associated degradation (ERAD), and mitochondrial protein import pathways.
In the HEK293T background, knockout of DNAJA2 disrupts these essential processes, compromising the cell’s ability to manage proteotoxic stress. This model is particularly relevant for studying diseases linked to protein misfolding and aggregation, including cancer and neurodegenerative disorders. The high protein synthesis capacity of HEK293T cells may exacerbate phenotypes associated with impaired protein quality control, providing a sensitized system for evaluating cellular responses to proteasome inhibition or other pharmacological challenges.
Researchers can employ this knockout cell population in a wide array of experimental contexts. Typical applications include dissecting the heat shock response, probing ER stress pathways, and validating DNAJA2 as a therapeutic target in cancers relying on proteostatic adaptation. Compatible assays encompass Western blotting for Hsp70 and stress markers, RT-qPCR of HSF1-regulated genes, aggregation and proteasome activity measurements, cell viability studies under stress, and co-immunoprecipitation of DNAJA2-Hsp70 complexes. For additional information or technical support, please contact Ascent Research.
