The DNAJA1 Knockout Caco-2 Cell Line is a CRISPR/Cas9-edited knockout cell line derived from the human colorectal adenocarcinoma Caco-2 cell line, with targeted disruption of the DNAJA1 gene leading to loss of its co-chaperone function. This model provides a defined system for investigating DNAJA1-dependent mechanisms in proteostasis within an intestinal epithelial background.
The parental Caco-2 cell line, established from a colon carcinoma, is widely used as an in vitro model of the intestinal epithelial barrier. Upon achieving confluence, Caco-2 cells spontaneously differentiate into a polarized, enterocyte-like monolayer, forming tight junctions and expressing brush border enzymes. This well-characterized system enables studies of intestinal biology, transport, and stress physiology.
DNAJA1, a member of the Hsp40 family, functions as a crucial co-chaperone that recruits Hsp70/Hsc70 to client proteins, facilitating their proper folding, trafficking, and degradation. Its expression is activated by HSF1 under stress conditions such as heat or oxidative challenge. DNAJA1 interacts with Hsp70, BAG family proteins (e.g., BAG3), and the E3 ligase CHIP to coordinate protein triage: refolding or proteasomal clearance. Disruption of DNAJA1 derails the Hsp70 chaperone cycle, causing accumulation of misfolded proteins and compromised cellular protein quality control.
In Caco-2 cells, DNAJA1 knockout imposes proteotoxic stress, mirroring pathologies where chaperone systems are overwhelmed, such as colorectal cancer and inflammatory bowel disease. The model enables dissection of how co-chaperone dysfunction influences epithelial barrier integrity, unfolded protein responses, and stress resilience. Given the constant environmental challenges faced by intestinal epithelia, this system is valuable for exploring how proteostasis disruption contributes to disease progression and intestinal homeostasis.
Key applications include profiling chaperone and stress gene expression by western blotting and RT-qPCR, measuring protein folding using luciferase-based reporters, and co-immunoprecipitation of Hsp70-client complexes. The cell line supports functional assays of cell viability under thermal or oxidative stress, proteasome activity, and autophagy flux analysis to evaluate aggregate clearance. It further serves as a screening platform for modulators of Hsp70?CDNAJA1 interactions, with potential implications in cancer and neurodegeneration therapeutics. For inquiries, contact Ascent Research.
