NAT-10 CRISPR Activation Plasmid (h): sc-406713-ACT

Human NAT10 encodes NAT-10, an acetyltransferase that modifies cytidine in RNA and acetylates protein substrates, supporting ribosome biogenesis, RNA metabolism, and nucleolar homeostasis. NAT-10 activity has been linked to N4-acetylcytidine (ac4C) deposition on mRNA and rRNA, influencing transcript stability and translational output, and it also participates in chromatin- and replication-associated processes that shape cell-cycle progression. Through these functions, NAT10 intersects with stress-response pathways and regulation of gene expression programs that govern proliferation and differentiation. Dysregulated NAT10 expression or activity has been reported across multiple disease contexts, including cancer-associated growth phenotypes and neurodevelopmental or premature aging–related cellular defects, making it a relevant node for mechanistic studies.

NAT-10 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NAT10 expression without altering the underlying DNA sequence.

NAT-10 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NAT10 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 NAT10 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NAT-10 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NAT10 locus and enabling the study of NAT-10-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NAT-10 pathway restoration in tumor cells with silenced or reduced NAT10 expression.

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