MAT IIα CRISPR Activation Plasmid (h): sc-402566-ACT

MAT2A encodes the catalytic alpha subunit of methionine adenosyltransferase II (MAT IIα), a key enzyme that generates S-adenosylmethionine (SAM), the principal methyl donor for DNA, RNA, and histone methylation as well as other methyltransferase-dependent reactions. By controlling SAM availability, MAT IIα links one-carbon metabolism and the methionine cycle to epigenetic regulation, chromatin state, and transcriptional programs that influence proliferation, differentiation, and stress responses. MAT2A activity is tightly coupled to nutrient status and cellular redox balance, integrating metabolic cues with methylation-dependent signaling. Dysregulation of MAT2A and SAM homeostasis has been associated with altered epigenetic landscapes and metabolic adaptation in disease-relevant contexts, making it a useful node for mechanistic studies of methylation and metabolic-epigenetic crosstalk.

MAT IIα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAT2A expression without altering the underlying DNA sequence.

MAT IIα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAT2A 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 MAT2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MAT IIα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAT2A locus and enabling the study of MAT IIα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MAT IIα pathway restoration in tumor cells with silenced or reduced MAT2A expression.

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