The DNAJB14 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population designed to disrupt the DNAJB14 gene, providing a robust loss-of-function model for investigating co-chaperone biology in a human colorectal adenocarcinoma background. This product consists of a heterogeneous pool of HT29 cells carrying targeted gene disruptions, enabling population-level studies without selection for single-cell clones. The CRISPR/Cas9-mediated editing introduces functional inactivation of DNAJB14, making this model suitable for examining consequences of impaired DnaJ/Hsp40 co-chaperone activity on cellular proteostasis, stress signaling, and cancer cell physiology.
The parental HT29 cell line is an aneuploid human colon carcinoma line with an epithelial morphology, widely employed in cancer research and intestinal epithelial biology. Derived from a primary colorectal adenocarcinoma, HT29 cells retain key features of transformed intestinal epithelium, including the ability to model barrier function, drug responses, and tumor-associated signaling. Their robust growth and well-characterized genetic background make them a standard host for gene disruption experiments exploring colorectal cancer pathology and therapeutic vulnerabilities.
DNAJB14 encodes a member of the DnaJ (Hsp40) family of co-chaperones that interacts directly with Hsp70 ATPases such as HSPA1A and HSPA8 to facilitate protein folding, translocation, and degradation. As part of the cellular quality control machinery, DNAJB14 participates in recognizing misfolded substrates and delivering them to Hsp70 for refolding or to the ubiquitin-proteasome system for clearance. Its function is transcriptionally regulated by HSF1 and is integrated with endoplasmic reticulum stress pathways via sensors PERK (EIF2AK3), IRE1 (ERN1), and ATF6, placing it at the intersection of cytoplasmic and ER protein homeostasis.
In HT29 colorectal cancer cells, disruption of DNAJB14 compromises Hsp70-mediated chaperone networks, leading to accumulation of misfolded proteins and induction of the unfolded protein response (UPR). This impairment alters sensitivity to proteotoxic stress and may affect cancer cell survival pathways reliant on efficient protein quality control. By perturbing interactions with co-chaperones such as BAG family members and Hsp70 isoforms, the knockout model highlights vulnerabilities linked to chaperone network integrity, offering insight into stress adaptation mechanisms exploited by colorectal carcinoma cells.
Researchers can use these polyclonal knockout cells to study chaperone function in intestinal cancer biology, including ER stress signaling, UPR dynamics, and sensitivity to agents such as proteasome inhibitors (e.g., bortezomib) or ER stress inducers (e.g., tunicamycin). Typical assays include western blotting for DNAJB14, Hsp70, and UPR markers like CHOP and BiP; RT-qPCR to assess XBP1 splicing; cell viability and apoptosis analyses via MTT or Annexin V/PI flow cytometry; and autophagy flux measurement through LC3 lipidation. Additionally, Transwell migration/invasion assays can evaluate the impact of disrupted proteostasis on cancer cell behavior. For further details or technical support, please contact Ascent Research.
