DNAJC16 Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from the Huh-7 cell line, featuring targeted disruption of the DNAJC16 gene. This polyclonal formulation provides a heterogeneous pool of edited cells, enabling robust loss-of-function studies without clonal selection artifacts. The cells are supplied as a ready-to-use mixed population for immediate experimental application.
Huh-7 is a well-differentiated hepatocellular carcinoma cell line that retains key hepatocyte features and is extensively used in liver cancer research. It supports the study of hepatic metabolism, viral infection, and drug response, making it an appropriate host for investigating genes implicated in liver disease.
DNAJC16 encodes an endoplasmic reticulum (ER) co-chaperone that directly interacts with BiP (HSPA5) to stimulate its ATPase activity, facilitating efficient protein folding and mitigating ER stress. It operates as a key node in the unfolded protein response (UPR) network, where it is activated by ER stress inducers such as tunicamycin and thapsigargin, and downstream of the sensor proteins ATF6, IRE1, and PERK. These sensors regulate transcription factors XBP1 and ATF4, which drive the expression of UPR target genes including CHOP (DDIT3). DNAJC16 associates with both BiP and GRP94 (HSP90B1), and interfaces with the ER-associated degradation (ERAD) machinery. Loss of DNAJC16 function disrupts BiP activity, leading to unresolved ER stress, accumulation of misfolded proteins, and a shift toward apoptotic signaling.
In Huh-7 hepatocellular carcinoma cells, DNAJC16 knockout creates a valuable model to dissect cancer cell reliance on ER proteostasis. It enables the study of how impaired co-chaperone function affects UPR signaling and cell survival, uncovering vulnerabilities that may be exploited for therapeutic intervention in liver cancer.
Researchers can utilize this knockout population to dissect UPR signaling through western blot analysis of BiP, CHOP, and phospho-eIF2??, as well as RT-qPCR for spliced XBP1. ER stress reporter assays, including luciferase-based systems, enable high-throughput screening of ER stress modulators. Co-immunoprecipitation assays with BiP and GRP94 explore co-chaperone dynamics, while immunofluorescence reveals alterations in ER structure. Cell viability assays following treatment with tunicamycin or thapsigargin quantify sensitivity to ER stress-induced death. These applications advance the understanding of hepatocellular carcinoma and facilitate the development of therapies targeting ER stress. For further details, please contact Ascent Research.
