The DIS3L2 Knockout HT29 Polyclonal Cells represent a CRISPR/Cas9-edited polyclonal knockout cell population derived from the human HT29 colorectal adenocarcinoma cell line, engineered to disrupt the DIS3L2 gene. This gene-edited population provides a loss-of-function model for studying the 3′-5′ exoribonuclease DIS3L2, a central mediator of terminal uridylation-dependent RNA decay. By abolishing DIS3L2 expression through CRISPR/Cas9-mediated gene disruption, researchers can systematically investigate the consequences of impaired degradation of uridylated RNA substrates on cellular physiology, transcriptome stability, and oncogenic signaling pathways.
The host HT29 cell line is an established epithelial model isolated from a female colorectal adenocarcinoma, characterized by mismatch repair proficiency and co-occurring BRAF V600E and p53 mutations. These genetic features render HT29 cells particularly relevant for dissecting colorectal cancer biology and intestinal epithelial differentiation. The cell line’s well-documented growth properties and signaling dependencies offer a robust platform for evaluating gene function in a disease-relevant genetic context, making it a widely adopted system for oncogene and tumor suppressor studies.
DIS3L2 functions as a critical surveillance enzyme that selectively degrades RNAs tagged by terminal uridylation, a process mediated by the TUTases TUT4 (ZCCHC11) and TUT7 (ZCCHC6). Through its exoribonuclease activity, DIS3L2 controls the levels of uridylated miRNAs, notably tumor-suppressive let-7 family members, and other uridylated transcripts. In the canonical pathway, LIN28 recruits TUT4/TUT7 to precursor let-7 (pre-let-7), promoting uridylation and subsequent degradation by DIS3L2, thereby fine-tuning mature let-7 expression. DIS3L2 additionally interacts with broader uridylated mRNA populations, linking its activity to post-transcriptional gene regulation. As a recognized tumor suppressor, its loss can lead to accumulation of oncogenic transcripts and aberrant miRNA profiles.
In the context of HT29 cells, deletion of DIS3L2 in a background harboring BRAF V600E and p53 mutations offers a unique opportunity to explore the interplay between RNA decay dysregulation and established colorectal cancer drivers. The knockout may result in elevated levels of uridylated let-7 precursors and other regulatory RNAs, potentially altering gene expression networks that control proliferation, apoptosis, and differentiation. This model enables mechanistic dissection of how loss of DIS3L2-mediated RNA surveillance contributes to miRNA imbalance and potentiates oncogenic signaling, providing insights into tumor suppression mechanisms operative in colorectal cancer and related malignancies.
This knockout cell population is ideally suited for a range of experimental applications, including functional genomics studies of RNA decay, miRNA turnover, and colorectal cancer pathobiology. Researchers can perform Western blotting to confirm DIS3L2 ablation, RT-qPCR and reporter assays to quantify let-7 activity and maturation, and RNA sequencing to map transcriptome-wide uridylation changes. Co-immunoprecipitation assays can be employed to probe DIS3L2 interactions with TUT4/TUT7, while cell-based assays for proliferation, apoptosis, colony formation, and xenograft tumor growth enable phenotypic characterization. The product also supports drug screening aimed at modulating RNA decay pathways and investigation of Perlman syndrome mechanisms. For further technical details and custom solutions, please contact Ascent Research.
