The DNAJB4 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal cell population generated from the HT29 human colon adenocarcinoma line, designed for loss-of-function studies of the DNAJB4 gene. This product delivers a heterogeneous pool of cells carrying CRISPR/Cas9-mediated gene disruptions, providing a population-level knockout model that avoids the potential biases of monoclonal isolation. The polyclonal format is well-suited for functional assays requiring robust knockout effects across a cell population.
HT29 cells are a widely employed human colorectal adenocarcinoma model derived from a primary tumor of a 44-year-old female patient. These adherent epithelial cells retain characteristics of colonic carcinoma, including the ability to form polarized monolayers and express cancer-relevant signaling pathways. The line is extensively used in colorectal cancer biology, intestinal epithelial barrier studies, and preclinical drug testing, making it an appropriate host for interrogating the tumor-suppressive functions of DNAJB4.
DNAJB4 is a member of the Hsp40 family and functions as a co-chaperone for Hsp70, facilitating protein folding, quality control, and proteostasis. It acts as a tumor suppressor, often downregulated in colorectal cancer. Upstream regulation involves heat shock, ER stress, and the transcription factor HSF1, as well as oncogenic Wnt and NF-??B signals. DNAJB4 interacts with Hsp70 and other Hsp40 co-chaperones to manage misfolded client proteins, influencing downstream targets such as Bcl-2 family members, the cell cycle inhibitor p21, cyclins, and matrix metalloproteinases. Knockout of DNAJB4 disrupts proteostatic control, leading to apoptosis resistance, activation of Wnt/??-catenin and NF-??B pathways, and increased proliferation and invasion.
In HT29 cells, loss of DNAJB4 mimics tumor-suppressor inactivation observed in colorectal tumorigenesis. The knockout cells exhibit compromised protein quality control and heightened oncogenic signaling, making them a valuable model for investigating the interplay between proteostasis and pathways such as Wnt/??-catenin and NF-??B. They also provide a platform for testing therapeutic agents targeting these networks and for studying epithelial barrier dysfunction associated with DNAJB4 loss.
This knockout cell model supports a broad range of applications including tumor suppressor mechanism studies, proteostasis in cancer, drug target validation, and functional genomics. Typical experiments involve western blotting to assess DNAJB4, Hsp70, and downstream targets; RT-qPCR for transcript quantification; MTT-based proliferation assays; Transwell migration and invasion tests; Annexin V apoptosis assays; co-immunoprecipitation to examine Hsp70 interactions; and RNA-seq for transcriptome-wide analysis. For additional technical details or ordering information, please contact Ascent Research.
