The DIS3L2 Knockout HAP1 Polyclonal Cells are a CRISPR/Cas9-edited population designed to disrupt the DIS3L2 gene in a human near-haploid background. This polyclonal pool contains a heterogeneous collection of edited alleles that collectively ablate DIS3L2 protein expression, avoiding clonal artifacts associated with single-cell isolation. The population is well-suited for loss-of-function studies and pooled genetic screens, providing a robust model to interrogate DIS3L2 function in RNA metabolism and disease.
HAP1 is a near-haploid human cell line derived from the KBM-7 chronic myeloid leukemia line. Its fibroblast-like morphology and stable near-haploid karyotype, disomic only for chromosome 8, simplify functional genomics by eliminating masking effects of heterozygous mutations. Widely adopted for CRISPR-based screens, HAP1 supports efficient editing and high-throughput assays owing to its male origin and adherent growth characteristics.
DIS3L2 encodes a 3??-5?? exoribonuclease that mediates decay of cytoplasmic RNAs bearing 3?? oligouridine tails, added by terminal uridylyl transferases TUT4 and TUT7 under regulation of Lin28A. DIS3L2 preferentially degrades structured substrates, including let-7 miRNA precursors and aberrant non-coding RNAs, functioning independently of the exosome in RNA surveillance. This pathway is central to developmental regulation; loss-of-function mutations cause Perlman syndrome, a congenital disorder with Wilms tumor predisposition and renal dysplasia.
In the near-haploid HAP1 context, DIS3L2 knockout provides a clean genetic system to dissect exosome-independent RNA decay. The absence of a second allele ensures unambiguous phenotypic attribution, facilitating analysis of the DIS3L2?CTUTase?Clet-7 axis. This model enables precise investigation of how DIS3L2 loss stabilizes oligouridylated transcripts and alters cell proliferation and apoptosis, mimicking Perlman syndrome?Cassociated pathophysiology.
Applications include RNA-seq and ribosome profiling to assess transcriptome-wide changes, RIP to characterize uridylated RNA substrates, and rescue experiments to confirm DIS3L2 interactions. RT-qPCR and reporter assays can quantify let-7 regulation, while proliferation and apoptosis assays evaluate functional outcomes. The polyclonal population is compatible with drug screening targeting RNA decay pathways. For further information, please contact Ascent Research.
