The DIS3L2 Knockout HGC-27 Polyclonal Cells constitute a CRISPR/Cas9-edited polyclonal knockout cell population in which the gene encoding the 3??-5?? exoribonuclease DIS3L2 has been disrupted. This loss-of-function model was generated using the HGC-27 gastric carcinoma cell line and is supplied as a heterogeneous pool of edited cells, allowing researchers to study the functional consequences of DIS3L2 ablation without clonal selection biases. The product enables investigation of DIS3L2-dependent RNA decay, microRNA biogenesis, and associated signaling pathways in a reproducible and accessible format.
HGC-27 is an epithelial cell line originally derived from a metastatic lymph node of a human gastric adenocarcinoma patient. This widely employed gastric cancer model exhibits characteristics relevant to advanced disease, including metastatic origin and maintained epithelial morphology. As a gastric adenocarcinoma line, HGC-27 provides a physiologically relevant context to explore molecular mechanisms underlying gastric cancer progression, enabling the dissection of oncogenic signaling, RNA regulation, and stress response pathways that are often dysregulated in gastrointestinal malignancies.
DIS3L2 functions as a cytoplasmic exoribonuclease that mediates exosome-independent degradation of uridylated RNA substrates. It is a key effector downstream of the PERK-ATF4 ER stress signaling axis, with ATF4 transcription factor directly upregulating DIS3L2 expression under stress conditions. DIS3L2 specifically recognizes and degrades RNAs marked with untemplated uridine tails added by the uridylyltransferases TUT4 (ZCCHC11) and TUT7 (ZCCHC6), and its activity is modulated by the RNA helicase MOV10. Known substrates include pre-let-7 microRNA precursors, whose uridylated forms are targeted for decay, as well as mRNAs encoding c-MYC, TP53, and apoptosis regulators, thereby linking DIS3L2 to the control of cell proliferation, apoptosis, and microRNA processing.
In the context of HGC-27 gastric cancer cells, knockout of DIS3L2 is predicted to disrupt the normal clearance of uridylated transcripts, leading to altered let-7 miRNA maturation and stabilization of mRNAs that influence cell growth and survival. Because let-7 miRNAs function as tumor suppressors that post-transcriptionally repress oncogenes, changes in let-7 levels due to impaired DIS3L2-mediated decay may amplify oncogenic signaling, making this model particularly relevant for gastric cancer research. Furthermore, perturbation of PERK-ATF4-dependent stress responses can be examined in a cell line that naturally exhibits cancer-associated stress phenotypes.
This polyclonal knockout cell population is suitable for a broad range of experimental applications. It supports gene expression analysis by RT-qPCR, protein detection via western blot, and transcriptome profiling by RNA-seq. Functional assays including MTS/MTT proliferation, Annexin V apoptosis, migration/invasion, and colony formation enable phenotypic characterization. The cells also serve as a platform for identifying uridylated RNA targets via 3??-RACE-seq, investigating let-7 microRNA regulation by qPCR, and screening for small molecules that modulate DIS3L2 activity. Investigators can additionally use RNA immunoprecipitation to probe DIS3L2?Csubstrate interactions or explore crosstalk with TUT4/TUT7 uridylyltransferases. For further information, please contact Ascent Research.
