DTX3L Knockout Huh-7 Polyclonal Cells are a CRISPR/Cas9-edited polyclonal knockout cell population derived from Huh-7 human hepatocellular carcinoma cells. This product features disruption of the DTX3L gene, encoding an E3 ubiquitin-protein ligase, and enables loss-of-function studies without clonal selection artifacts. The knockout model is suitable for investigating DTX3L-dependent ubiquitin signaling in DNA damage response and interferon pathways.
Huh-7 cells are a well-differentiated human hepatocellular carcinoma line retaining liver parenchymal functions, including metabolic, synthetic, and detoxification activities. These cells are extensively used for hepatitis C virus replication studies and liver cancer research, providing a physiologically relevant system for hepatocyte biology. Their hepatocellular origin makes them ideal for examining the roles of ubiquitin ligases like DTX3L in hepatocarcinogenesis and antiviral innate immunity.
DTX3L functions as an E3 ubiquitin ligase that, in complex with PARP9, catalyzes ubiquitination of histone H2B, modulating chromatin structure and gene expression during DNA damage repair and interferon signaling. Upstream, DTX3L is activated by interferon alpha/gamma (IFNA/IFNG) and DNA damage signals, leading to recruitment to chromatin. Downstream targets include histone H2A and transcription factors STAT1/STAT2. It interacts with PARP9 and ubiquitin-conjugating enzymes, and feeds into the RIG-I-like receptor pathway through effectors such as RIG-I, MAVS, TBK1, IRF3, and IRF7. Knockout abrogates histone H2B ubiquitination, potentially impairing DNA repair and attenuating interferon-stimulated gene expression.
In Huh-7 cells, DTX3L knockout offers a model to dissect ubiquitin-mediated regulation at the intersection of DNA damage repair and antiviral signaling. Hepatocytes face genotoxic and viral challenges; loss of DTX3L may reveal vulnerabilities in liver cancer cells, as DTX3L is implicated in hepatocellular carcinoma and interferon responses. This model can be used to study how impaired chromatin modification affects tumor cell survival and immune evasion, and may sensitize cells to DNA-damaging agents or disrupt innate immune responses.
Applications include Western blotting for histone H2B ubiquitination, co-immunoprecipitation of DTX3L-PARP9 complexes, RT-qPCR for interferon-stimulated genes, ??H2AX immunofluorescence, comet assays, RNA-seq transcriptome profiling, flow cytometry for STAT1 phosphorylation, and MTT viability assays. This knockout model is suitable for target validation in liver cancer, functional studies of ubiquitin signaling, and host-pathogen interaction research. For more information, please contact Ascent Research.
