CPS1 CRISPR Activation Plasmid (h): sc-402014-ACT

Human CPS1 encodes carbamoyl-phosphate synthase 1, a mitochondrial enzyme that catalyzes the first committed step of the urea cycle by converting ammonia and bicarbonate into carbamoyl phosphate using ATP. This activity is central to hepatic nitrogen disposal and intersects with amino acid catabolism, mitochondrial metabolism, and downstream arginine biosynthesis. Altered CPS1 expression or function is linked to hyperammonemia and broader metabolic dysregulation, and CPS1 status is frequently evaluated in studies of liver physiology, mitochondrial stress responses, and nitrogen balance. In cancer and metabolic research, CPS1 is also used as a context-dependent marker of metabolic reprogramming and lineage-associated urea cycle activity.

CPS1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CPS1 expression without altering the underlying DNA sequence.

CPS1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CPS1 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 CPS1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CPS1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CPS1 locus and enabling the study of CPS1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CPS1 pathway restoration in tumor cells with silenced or reduced CPS1 expression.

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