PARG CRISPR Activation Plasmid (h): sc-404382-ACT

Poly(ADP-ribose) glycohydrolase (PARG) is the primary enzyme responsible for hydrolyzing poly(ADP-ribose) (PAR) chains synthesized by PARP family members, thereby terminating PARylation signaling and restoring protein function after stress responses. By regulating PAR turnover, PARG helps control DNA damage sensing and repair, chromatin remodeling, replication stress tolerance, and cell death pathways such as parthanatos through modulation of PAR-dependent protein recruitment. PARG activity interfaces with base excision repair and single-strand break repair processes, influencing genome stability and transcriptional programs linked to cellular stress adaptation. Dysregulated PAR metabolism has been associated with cancer biology and neurodegeneration, making PARG a useful node for mechanistic studies of DNA repair capacity, chromatin dynamics, and stress-induced signaling.

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

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

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