NQO2 CRISPR Activation Plasmid (h): sc-402200-ACT

Human NQO2 (NRH:quinone oxidoreductase 2) encodes a cytosolic flavoprotein that catalyzes two-electron reduction of quinones and related electrophiles, influencing cellular redox balance and detoxification capacity. By modulating quinone cycling and reactive oxygen species formation, NQO2 intersects with oxidative stress responses, xenobiotic metabolism, and broader NAD(P)H-dependent redox networks. Altered NQO2 expression and activity have been associated with phenotypes linked to metabolic stress, inflammation, and cellular susceptibility to electrophile-mediated damage, making it relevant for mechanistic studies of stress adaptation. In cancer biology and neurobiology research, NQO2 is often examined in the context of redox homeostasis, DNA damage sensitivity, and signaling changes driven by electrophilic metabolites.

NQO2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NQO2 expression without altering the underlying DNA sequence.

NQO2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NQO2 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 NQO2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NQO2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NQO2 locus and enabling the study of NQO2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NQO2 pathway restoration in tumor cells with silenced or reduced NQO2 expression.

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