BMP-9 CRISPR Activation Plasmid (h): sc-402887-ACT

GDF2 encodes bone morphogenetic protein 9 (BMP-9), a secreted TGF-β superfamily ligand that signals primarily through type I receptors such as ALK1 and type II receptors including BMPR2 to activate SMAD1/5/9-dependent transcription. In human cells, BMP-9 helps regulate endothelial quiescence, vascular remodeling, and angiogenic balance, and also influences osteogenic and metabolic gene programs in a context-dependent manner. BMP-9 pathway activity intersects with BMP/TGF-β crosstalk, extracellular matrix regulation, and inflammatory signaling, shaping cell fate decisions across vascular, hepatic, and mesenchymal compartments. Dysregulated BMP-9/GDF2 signaling has been associated in the literature with vascular pathobiology, including heritable and acquired disorders of endothelial function and remodeling, supporting its relevance as a research target in cardiovascular and developmental biology.

BMP-9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GDF2 expression without altering the underlying DNA sequence.

BMP-9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GDF2 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 GDF2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BMP-9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GDF2 locus and enabling the study of BMP-9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BMP-9 pathway restoration in tumor cells with silenced or reduced GDF2 expression.

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