The DIS3L Knockout Huh-7 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated from the Huh-7 human hepatocellular carcinoma cell line. This product is designed for loss-of-function studies of the DIS3L gene, which encodes a critical catalytic subunit of the cytoplasmic RNA exosome. The polyclonal pool consists of a heterogeneous mixture of cells with targeted gene disruptions, enabling robust investigation of DIS3L function in cytoplasmic RNA degradation and surveillance without the need for monoclonal selection.
The Huh-7 cell line was originally derived from a hepatocellular carcinoma of a 57-year-old Japanese male and is extensively used as a model for liver cancer research. These epithelial cells retain hepatocellular features and support studies of hepatic gene expression, signal transduction, and viral replication. The line??s genetic background provides a physiologically relevant platform for examining molecular pathways involved in hepatocarcinogenesis and RNA metabolism.
DIS3L furnishes the 3′-5′ exonuclease activity of the cytoplasmic exosome complex, which degrades a broad range of substrates including cytoplasmic mRNAs, microRNAs, and long non-coding RNAs. It operates in association with the exosome core components EXOSC2 through EXOSC10 and collaborates with the SKI complex (SKI2 and SKIV2L) and the nonsense-mediated decay factor UPF1 to recognize and unwind RNA targets. DIS3L expression is controlled by the MYC transcription factor and cellular stress signals. Loss of DIS3L disrupts cytoplasmic RNA surveillance, leading to the accumulation of aberrant transcripts that perturb downstream gene networks and cellular homeostasis.
Within the Huh-7 hepatocellular carcinoma background, DIS3L ablation creates a powerful system for exploring the intersection of RNA decay defects and liver cancer. The buildup of exosome substrates can mimic RNA processing stress associated with hepatocarcinogenesis, impacting cell proliferation, apoptosis, and drug sensitivity. This polyclonal knockout model is particularly well-suited for identifying specific RNAs whose dysregulation drives malignant phenotypes and for assessing the therapeutic potential of targeting RNA surveillance pathways in HCC.
Key research applications include transcriptome-wide RNA-seq to map DIS3L-dependent RNA substrates, metabolic labeling-based RNA stability assays, and co-immunoprecipitation to profile exosome complex remodeling upon DIS3L loss. Functional assays for cell proliferation, apoptosis, and migration link DIS3L deficiency to cancer-relevant phenotypes. Western blotting and RT-qPCR confirm knockout efficiency and quantify changes in target RNA levels, while immunofluorescence reveals alterations in exosome localization. These approaches enable detailed dissection of RNA surveillance mechanisms and may uncover novel therapeutic vulnerabilities in hepatocellular carcinoma. For further details or to discuss custom applications, please contact Ascent Research.
