ARH1 CRISPR Activation Plasmid (h2): sc-410678-ACT-2

Human ADPRH encodes ADP-ribosylarginine hydrolase 1 (ARH1), a cytosolic enzyme that removes mono-ADP-ribose from arginine residues on proteins, reversing arginine-specific ADP-ribosylation and shaping cellular ADP-ribose turnover. By counterbalancing ART-mediated modification, ARH1 contributes to regulation of stress responses, signal transduction, and protein homeostasis within broader NAD⁺-dependent mono-ADP-ribosylation pathways. Dysregulated ADP-ribosylation dynamics involving ADPRH have been linked to altered susceptibility to toxin-mediated modifications and to phenotypes relevant to tumor suppression and inflammatory signaling in experimental systems. Gene editing of ADPRH enables mechanistic studies of reversible ADP-ribosylation, pathway cross-talk with DNA damage and immune signaling networks, and functional genomics screens examining post-translational modification control.

ARH1 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous ADPRH expression without altering the underlying DNA sequence.

ARH1 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADPRH 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 ADPRH transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARH1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADPRH locus and enabling the study of ARH1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARH1 pathway restoration in tumor cells with silenced or reduced ADPRH expression.

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