NAT-8 CRISPR Activation Plasmid (h): sc-411236-ACT

Human NAT8 encodes NAT-8, a kidney- and liver-enriched N-acetyltransferase localized primarily to endoplasmic reticulum membranes that catalyzes N-acetylation reactions impacting small-molecule and xenobiotic handling. NAT-8 activity is linked to metabolic homeostasis and cellular detoxification processes, intersecting with pathways that influence oxidative stress responses and tubular epithelial function. Genetic variation and altered expression of NAT8 have been associated with renal traits and susceptibility to kidney injury, making it a useful target for mechanistic studies of nephrotoxicity and metabolic stress. In cell models, NAT-8 expression can modulate phenotypes related to epithelial transport, cellular resilience to chemical exposure, and injury-associated transcriptional programs.

NAT-8 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NAT8 expression without altering the underlying DNA sequence.

NAT-8 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NAT8 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 NAT8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NAT-8 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NAT8 locus and enabling the study of NAT-8-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NAT-8 pathway restoration in tumor cells with silenced or reduced NAT8 expression.

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