NQO1 CRISPR Activation Plasmid (m): sc-421913-ACT

Mouse Nqo1 encodes NAD(P)H quinone dehydrogenase 1 (NQO1), a cytosolic flavoprotein that catalyzes two-electron reduction of quinones to hydroquinones, limiting redox cycling and reactive oxygen species generation. NQO1 functions downstream of oxidative and electrophilic stress signaling and is commonly regulated within NRF2-driven antioxidant response and xenobiotic metabolism programs. By shaping cellular redox homeostasis, detoxification capacity, and proteostasis, NQO1 influences susceptibility to oxidative injury and metabolic stress phenotypes. Altered Nqo1/NQO1 activity is frequently examined in models of carcinogenesis, inflammation, neurodegeneration, and cardiometabolic dysfunction where redox imbalance and electrophile handling are central mechanisms.

NQO1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nqo1 expression without altering the underlying DNA sequence.

NQO1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nqo1 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 Nqo1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NQO1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nqo1 locus and enabling the study of NQO1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NQO1 pathway restoration in tumor cells with silenced or reduced Nqo1 expression.

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