MGP CRISPR Activation Plasmid (h): sc-401556-ACT

Human MGP (matrix Gla protein) is a secreted, vitamin K–dependent extracellular matrix protein best known for inhibiting ectopic mineralization by binding calcium and regulating hydroxyapatite deposition. It is highly relevant to vascular biology, where it modulates osteogenic signaling programs in vascular smooth muscle cells and interfaces with BMP/TGF-β-related pathways that govern calcification, remodeling, and extracellular matrix homeostasis. Dysregulated MGP expression or carboxylation status is associated with pathological calcification phenotypes observed in cardiovascular and connective tissue contexts, making it a useful molecular handle for studying calcification control mechanisms. As a circulating and matrix-associated factor, MGP is also leveraged as a biomarker-linked gene in mechanistic studies of mineral metabolism and tissue aging.

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

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

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