MTAP CRISPR Activation Plasmid (h): sc-406223-ACT

Human MTAP (methylthioadenosine phosphorylase) catalyzes phosphorolysis of 5′-methylthioadenosine to adenine and 5-methylthioribose-1-phosphate, linking the methionine salvage pathway to purine metabolism and cellular methylation balance. By maintaining adenine supply and regulating MTA levels, MTAP influences polyamine metabolism, transmethylation flux, and broader one‑carbon–related biochemical homeostasis. MTAP loss is frequently observed in cancers with 9p21 deletions near CDKN2A/CDKN2B, creating characteristic metabolic dependencies and altered signaling contexts relevant to tumor biology. Modulating MTAP expression is therefore useful for dissecting metabolic rewiring, nucleotide homeostasis, and epigenetic cofactor availability in disease-relevant models.

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

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

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