DIS3L2 CRISPR Activation Plasmid (h): sc-405372-ACT

Human DIS3L2 encodes a cytoplasmic 3′–5′ exoribonuclease that participates in RNA surveillance and turnover, with a prominent role in degrading oligouridylated RNA substrates produced by TUTase-mediated tailing. Through selective clearance of aberrant pre-miRNAs and structured RNAs, DIS3L2 influences small RNA biogenesis, post-transcriptional gene regulation, and cellular homeostasis under stress. DIS3L2 activity interfaces with pathways controlling proliferation and differentiation by shaping transcript stability and RNA quality control networks. Altered DIS3L2 function has been linked to developmental and tumor-associated phenotypes, making it a useful node for studying how dysregulated RNA decay contributes to disease-relevant gene expression programs.

DIS3L2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous DIS3L2 expression without altering the underlying DNA sequence.

DIS3L2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the DIS3L2 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the DIS3L2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DIS3L2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native DIS3L2 locus and enabling the study of DIS3L2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DIS3L2 pathway restoration in tumor cells with silenced or reduced DIS3L2 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.