ARH2 CRISPR Activation Plasmid (m): sc-433335-ACT

Mouse Adprhl1 encodes ARH2, a member of the ADP-ribosylhydrolase-like family implicated in regulating mono-ADP-ribosylation dynamics and related protein turnover cues. Although enzymatic activity for some family members can be atypical, ARH2 is studied for its contributions to cellular stress responses, metabolic control, and signaling processes that intersect with DNA damage handling and redox homeostasis. In cardiac and muscle biology models, Adprhl1 expression has been linked to myofibrillar organization and developmental programs, making it relevant to investigations of cardiomyopathy-associated pathways and tissue remodeling mechanisms. These properties position ARH2 as a useful node for dissecting how ADP-ribose–associated signaling interfaces with organ-specific physiology.

ARH2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Adprhl1 expression without altering the underlying DNA sequence.

ARH2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Adprhl1 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 Adprhl1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ARH2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Adprhl1 locus and enabling the study of ARH2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ARH2 pathway restoration in tumor cells with silenced or reduced Adprhl1 expression.

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