ME1 CRISPR Activation Plasmid (h): sc-401587-ACT

Human ME1 encodes cytosolic NADP-dependent malic enzyme 1, which catalyzes conversion of malate to pyruvate while generating NADPH for reductive biosynthesis and antioxidant defense. By supplying NADPH, ME1 supports fatty acid and cholesterol synthesis, maintains redox balance through glutathione recycling, and links glycolytic and TCA-cycle intermediates to anabolic metabolism. ME1 activity intersects with central carbon metabolism and NADPH-dependent pathways that influence cellular proliferation, oxidative stress responses, and differentiation programs. Altered ME1 expression has been associated with metabolic rewiring observed in diverse disease-relevant contexts, including cancer metabolism, insulin resistance, and lipid dysregulation, making it a useful node for mechanistic studies.

ME1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ME1 expression without altering the underlying DNA sequence.

ME1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ME1 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 ME1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ME1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ME1 locus and enabling the study of ME1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ME1 pathway restoration in tumor cells with silenced or reduced ME1 expression.

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