The HDDC3 Knouckout HT29 Polyclonal Cells product comprises a CRISPR/Cas9-edited polyclonal knockout cell population targeting the HDDC3 gene in the HT29 human colorectal adenocarcinoma cell line. This gene-disrupted model provides a powerful tool for loss-of-function studies of HDDC3, an interferon-stimulated phosphohydrolase implicated in antiviral innate immunity. The polyclonal format ensures a diverse allelic knockout pool, enabling robust functional analyses without clonal selection artifacts. Researchers can exploit this cell population to interrogate HDDC3-dependent mechanisms in a well-characterized epithelial background relevant to intestinal biology and oncology.
HT29 cells are derived from a human colorectal adenocarcinoma and represent a widely used intestinal epithelial model. They harbor mutations in APC and TP53, alongside a BRAF V600E mutation, while maintaining wild-type KRAS, making them a cornerstone for colorectal cancer research and drug response studies. This cell line retains key characteristics of enterocytes, including the capacity to mount interferon responses, and thus provides a physiologically relevant system for exploring innate immune pathways. The integration of HDDC3 knockout into this genetic context enables dissection of gene function within a defined oncogenic background.
HDDC3 encodes a phosphohydrolase strongly induced by type I interferons (IFN-??/??) through the JAK-STAT cascade. IFNAR1 binding activates JAK1 and STAT1, forming the ISGF3 complex with IRF9 to transactivate genes via ISRE promoters, including HDDC3. The protein interacts with HCV NS5A and NS4B, disrupting the viral RNA replication complex. Upstream viral RNA sensors RIG-I, MDA5, and TLR3 initiate this pathway, positioning HDDC3 as a downstream antiviral effector. Beyond HCV, HDDC3 may contribute to broader ISG-mediated immunity.
The HT29 colorectal adenocarcinoma model is particularly valuable for studying the intersection of innate immunity and oncogenesis. Intestinal epithelial cells are frontline responders to viral pathogens, and HDDC3’s role in restricting HCV RNA replication suggests analogous functions against enteric viruses. Moreover, given the constitutive interferon signaling often observed in cancer, this knockout model facilitates examination of how HDDC3 influences cellular processes such as proliferation, apoptosis, and response to immunomodulatory therapies. The presence of APC and TP53 mutations further allows for investigation of potential crosstalk between tumor suppressor pathways and HDDC3-mediated antiviral mechanisms.
Key applications include interferon stimulation assays, co-immunoprecipitation with viral proteins, and viral replication assays using HCV replicons or enteric viruses. The model suits CRISPR screens for anticancer ISGs and transcriptomic profiling via RNA-seq. Cell proliferation and apoptosis assays further dissect HDDC3’s role in cancer biology. Target disruption is confirmed by Western blotting and RT-qPCR. For further details, please contact Ascent Research.
