NUDT21 CRISPR Activation Plasmid (h): sc-402082-ACT

Human NUDT21 encodes a core component of the cleavage factor Im (CFIm) complex that recognizes UGUA elements on pre-mRNA and promotes 3′ end processing and poly(A) site choice. By regulating alternative polyadenylation, NUDT21 shapes 3′ UTR length, transcript stability, localization, and translational control, thereby influencing global gene expression programs. This RNA-processing role links NUDT21 to pathways governing cell-cycle control, differentiation, and stress responses through coordinated modulation of mRNA isoform usage. Dysregulated CFIm activity and altered alternative polyadenylation patterns involving NUDT21 have been associated with disease-relevant transcriptional remodeling in cancer and neurodevelopmental phenotypes, making it a useful node for mechanistic studies of post-transcriptional regulation.

NUDT21 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NUDT21 expression without altering the underlying DNA sequence.

NUDT21 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NUDT21 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 NUDT21 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NUDT21 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NUDT21 locus and enabling the study of NUDT21-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NUDT21 pathway restoration in tumor cells with silenced or reduced NUDT21 expression.

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