ME2 CRISPR Activation Plasmid (h): sc-402900-ACT

Human ME2 encodes the mitochondrial NAD-dependent malic enzyme that catalyzes oxidative decarboxylation of malate to pyruvate, linking the tricarboxylic acid cycle with mitochondrial redox balance and anaplerotic/cataplerotic flux. By producing NADH and regulating pyruvate availability, ME2 supports oxidative metabolism, reactive oxygen species homeostasis, and biosynthetic programs coupled to proliferative states. Altered ME2 activity has been associated with metabolic reprogramming and mitochondrial dysfunction phenotypes observed across cancer biology and neurodegeneration research, where shifts in malate–pyruvate interconversion influence energetic and redox constraints. ME2 is therefore widely studied in pathways governing mitochondrial metabolism, redox signaling, and stress responses.

ME2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ME2 expression without altering the underlying DNA sequence.

ME2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ME2 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 ME2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ME2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ME2 locus and enabling the study of ME2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ME2 pathway restoration in tumor cells with silenced or reduced ME2 expression.

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