MetAP-2 CRISPR Activation Plasmid (h): sc-402529-ACT

Human METAP2 encodes methionine aminopeptidase 2 (MetAP-2), a cytosolic metalloprotease that removes the initiator methionine from nascent polypeptides, a key step in N-terminal processing that can influence protein stability, localization, and downstream N-acetylation. Through its role in co-translational protein maturation, MetAP-2 supports proteostasis and impacts translation-coupled growth programs and cell-cycle progression. METAP2 activity has been linked to pathways governing proliferative capacity and cellular stress adaptation, making it relevant to mechanistic studies of dysregulated growth in oncology and to investigations of metabolic and angiogenic phenotypes. Altered METAP2 expression or function can therefore serve as a node for interrogating how N-terminal processing interfaces with signaling networks and cellular fitness.

MetAP-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous METAP2 expression without altering the underlying DNA sequence.

MetAP-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the METAP2 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 METAP2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous MetAP-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native METAP2 locus and enabling the study of MetAP-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of MetAP-2 pathway restoration in tumor cells with silenced or reduced METAP2 expression.

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