BMP-1 CRISPR Activation Plasmid (h): sc-402432-ACT

BMP1 encodes bone morphogenetic protein 1 (BMP-1), a secreted metalloprotease of the astacin family that drives extracellular matrix (ECM) maturation by proteolytically processing procollagens and activating matrix-associated factors. Through cleavage events that support collagen fibrillogenesis and basement membrane organization, BMP-1 contributes to connective tissue architecture, tissue remodeling, and wound repair–associated ECM turnover. BMP-1 activity functionally intersects with TGF-β signaling and matricellular pathways by modulating the availability and activation state of ECM-bound growth factors and structural components. Dysregulated BMP1 expression or protease activity has been implicated in fibrotic remodeling and heritable connective tissue phenotypes characterized by altered collagen processing.

BMP-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BMP1 expression without altering the underlying DNA sequence.

BMP-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BMP1 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 BMP1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BMP-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BMP1 locus and enabling the study of BMP-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BMP-1 pathway restoration in tumor cells with silenced or reduced BMP1 expression.

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