The DIS3L2 Knockout HEK293T Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population generated by CRISPR/Cas9-mediated disruption of the DIS3L2 gene in HEK293T cells. This product provides a heterogeneous pool of cells with target-gene disruption, enabling loss-of-function studies without clonal selection. The polyclonal format preserves genetic diversity while enriching for DIS3L2-deficient cells, making it suitable for bulk biochemical and functional assays. DIS3L2 encodes a 3′-5′ exoribonuclease critical for cytoplasmic RNA turnover, and its inactivation is central to investigating RNA decay pathways and associated diseases.
The host cell line, HEK293T, is a widely used derivative of human embryonic kidney 293 cells, transformed with adenovirus type 5 DNA and stably expressing the SV40 large T antigen. This genetic background supports high-level protein expression and efficient viral production, positioning HEK293T as a workhorse for cell biology research, including transient transfection and lentiviral packaging. The cells?? robust growth and ease of manipulation facilitate reproducible knockout experiments. The integration of the DIS3L2 disruption into this well-characterized host provides a versatile platform for studying RNA metabolism in a human cellular context.
DIS3L2 functions as a key 3′-5′ exoribonuclease in cytoplasmic RNA surveillance, specifically recognizing and degrading uridylated RNA species. Its activity is tightly linked to the RNA uridylation machinery, including the terminal uridylyltransferases TUT4 and TUT7, which add uridine tails to target RNAs, marking them for DIS3L2-mediated decay. Major downstream targets include miRNAs of the let-7 family and oncogenic mRNAs, whose stability is modulated by uridylation status. DIS3L2 interacts with TUT4, TUT7, and RNA helicases to form a coordinated degradation complex. This pathway, together with the exosome complex (EXOSC), constitutes a critical node in post-transcriptional gene regulation. Mutations in DIS3L2 are linked to Perlman syndrome, Wilms tumor, and other overgrowth syndromes, underscoring its role in developmental and cancer biology.
In the HEK293T background, DIS3L2 knockout leads to accumulation of uridylated RNAs, mirroring molecular phenotypes observed in disease states. The high-expression capacity of HEK293T cells amplifies these effects, facilitating detection and characterization of stabilized RNA species. This model enables detailed dissection of DIS3L2-dependent RNA decay mechanisms and their impact on gene expression networks. The polyclonal nature avoids artifacts from clonal variation while providing a robust system for evaluating global RNA changes. Researchers can leverage this system to explore how loss of DIS3L2 influences oncogenic pathways or developmental signaling, given the cancer-associated role of its targets.
This polyclonal knockout cell population is suitable for diverse research applications, including mechanistic studies of RNA degradation, miRNA regulation, and uridylation-mediated decay. Typical experimental approaches include Western blotting to confirm DIS3L2 protein loss, RT-qPCR to quantify let-7 miRNA levels, and RNA sequencing to profile uridylated RNA accumulation. Functional assays such as cell proliferation and apoptosis assays can link DIS3L2 loss to cellular phenotypes. The model further supports drug discovery screens targeting RNA decay pathways or RNA-modifying enzymes. For further information or technical support, please contact Ascent Research.
