The DIS3L2 Knockout Huh-7 Polyclonal Cells consist of a population of Huh-7 human hepatocellular carcinoma cells subjected to CRISPR/Cas9-mediated disruption of the DIS3L2 gene. This product is supplied as a heterogeneous pool of edited cells, avoiding clonal artifacts and faithfully representing the complexity of gene knockout in a polyclonal format. The model enables investigation of DIS3L2-dependent RNA decay and tumor suppressor functions without single-cell-derived bias, making it suitable for high-content functional genomic studies.
Huh-7 cells are an epithelial cell line originating from the liver tumor of a 57-year-old Japanese male. This hepatocellular carcinoma model retains hepatocyte features and is permissive for hepatitis C virus replication, rendering it a widely adopted platform for liver cancer research and hepatic pathogenesis. The line??s established molecular profile and responsiveness to oncogenic stimuli provide a robust background for targeted gene disruption.
DIS3L2 encodes a 3′-5′ exoribonuclease that degrades uridylated RNA substrates, acting as a key effector in RNA surveillance and microRNA biogenesis. The enzyme functions downstream of LIN28A/B and terminal uridylyltransferases TUT4/TUT7, which uridylate pre-let-7 transcripts to mark them for DIS3L2-mediated decay. Consequently, DIS3L2 loss leads to accumulation of uridylated pre-let-7, suppression of mature let-7, and stabilization of oncogenic mRNAs including LIN28B, c-MYC, CCND1, and IGF2. This disruption of the LIN28A-TUT4-pre-let-7-DIS3L2-let-7 axis drives dysregulation of cell proliferation and survival pathways. DIS3L2 is a recognized tumor suppressor, and its deficiency is associated with Perlman syndrome and Wilms tumor.
In the context of hepatocellular carcinoma, DIS3L2 knockout in Huh-7 cells provides insight into how impaired RNA decay contributes to liver tumorigenesis. The model highlights the interplay between the LIN28/let-7 pathway and hepatic transformation, offering a system to probe mechanisms by which DIS3L2 loss exacerbates malignant phenotypes such as uncontrolled growth and apoptosis resistance. The HCV-permissive nature of Huh-7 further enables studies on viral interactions with host RNA decay machinery, potentially revealing new facets of virus-induced liver pathology.
Typical experimental applications include RT-qPCR analysis of let-7 levels, RNA sequencing to identify accumulated uridylated transcripts, and western blotting for downstream effectors like c-MYC and CCND1. Functional assays such as cell proliferation, colony formation, and apoptosis measurements assess tumor-suppressive impacts. Co-immunoprecipitation experiments with TUT4 or TUT7 can examine enzyme-substrate dynamics. Additionally, these cells are well-suited for drug screens targeting the LIN28/let-7/DIS3L2 pathway or for identifying synthetic lethal interactions in liver cancer. For further information, please contact Ascent Research.
