NUDT8 CRISPR Activation Plasmid (m): sc-425987-ACT

Mouse Nudt8 encodes NUDT8, a member of the Nudix hydrolase family that catalyzes hydrolysis of nucleoside diphosphate derivatives to maintain nucleotide pool quality and limit accumulation of potentially toxic metabolites. By controlling levels of oxidized or aberrant nucleotide species, NUDT8 contributes to metabolic homeostasis and helps protect nucleic acids from misincorporation and damage. Nudix enzymes are commonly linked to cellular stress responses, redox balance, and DNA/RNA integrity, making Nudt8 expression and regulation relevant to studies of genome stability and inflammation-associated signaling. Altered nucleotide sanitation pathways are frequently investigated in the context of cancer biology, neurodegeneration, and metabolic dysregulation, where changes in nucleotide turnover can influence mutational burden and cell survival programs.

NUDT8 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nudt8 expression without altering the underlying DNA sequence.

NUDT8 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nudt8 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 Nudt8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NUDT8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nudt8 locus and enabling the study of NUDT8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NUDT8 pathway restoration in tumor cells with silenced or reduced Nudt8 expression.

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