ARG1/Arginase 1 CRISPR Activation Plasmid (h): sc-418076-ACT

Human ARG1 encodes arginase 1, a cytosolic metalloenzyme that catalyzes conversion of L-arginine to L-ornithine and urea, serving as a terminal step of the urea cycle and a key regulator of nitrogen disposal. By controlling intracellular arginine availability, ARG1 influences polyamine and proline biosynthesis through ornithine flux and can modulate nitric oxide synthesis indirectly via competition with nitric oxide synthases. ARG1 activity is closely linked to hepatocyte metabolic homeostasis and to immunometabolic programs in myeloid cells, where arginine depletion can shape T cell responsiveness and inflammatory signaling. Dysregulated ARG1 expression or function is associated with disorders of ammonia detoxification and has been implicated in tumor-associated myeloid biology and other contexts where arginine metabolism is remodeled.

ARG1/Arginase 1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ARG1 expression without altering the underlying DNA sequence.

ARG1/Arginase 1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ARG1 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 ARG1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARG1/Arginase 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ARG1 locus and enabling the study of ARG1/Arginase 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARG1/Arginase 1 pathway restoration in tumor cells with silenced or reduced ARG1 expression.

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