This product consists of a CRISPR/Cas9-edited polyclonal knockout population of HEK293T cells specifically targeting the DIS3L gene. The polyclonal format provides a heterogeneous pool of edited cells, enabling researchers to study the consequences of DIS3L loss in a representative cellular context without single-cell clonal isolation. The editing strategy disrupts the DIS3L locus, establishing a loss-of-function model suitable for functional genomics, RNA biology, and mechanistic investigations.
The HEK293T host cell line is derived from human embryonic kidney epithelial cells and is widely utilized in biomedical research for its robust protein expression, efficient viral production, and tractable growth characteristics. The cells stably express the SV40 large T antigen, which supports episomal replication of plasmids containing the SV40 origin. These attributes make HEK293T an ideal platform for exploring gene function in a human cell background, particularly for genes involved in fundamental processes such as RNA metabolism and gene regulation.
DIS3L encodes a catalytically inactive subunit of the RNA exosome complex, serving as a scaffold protein that organizes the core exosome structure and facilitates the recruitment of active exoribonucleases like DIS3 and DIS3L2. Functioning within multiple RNA decay pathways??including mRNA surveillance, nonsense-mediated decay, and AU-rich element (ARE)-mediated degradation??DIS3L interacts with EXOSC2, EXOSC3, and SKI complex subunits to regulate the turnover of specific mRNAs. Its activity is modulated by cellular stress signals and ARE-binding factors, and its disruption leads to accumulation of transcripts encoding cell cycle regulators and oncoproteins, thereby perturbing gene expression homeostasis.
In the HEK293T context, knockout of DIS3L impairs the structural integrity of the RNA exosome, thereby providing a powerful model to study exosome-mediated RNA decay mechanisms. The human embryonic kidney origin and the well-characterized transcriptional landscape of HEK293T cells enable precise dissection of how loss of this scaffold protein affects global RNA stability, processing, and surveillance pathways. This model is particularly valuable for investigating the interplay between the exosome and other decay machineries, such as the CCR4-NOT deadenylase complex, in a genetically accessible system.
Researchers can employ this DIS3L knockout polyclonal population to assess changes in transcriptome-wide RNA abundance via RNA-seq, validate specific mRNA targets by RT-qPCR or Northern blotting, and monitor protein expression of downstream effectors through western blotting. Functional studies may include proliferation and apoptosis assays to explore the cellular consequences of altered RNA homeostasis, with potential implications for understanding the role of exosome dysfunction in cancer biology. For additional information, pricing, or technical inquiries, please contact Ascent Research.
