BMPR-II CRISPR Activation Plasmid (h): sc-400895-ACT

BMPR2 encodes bone morphogenetic protein receptor type II (BMPR-II), a serine/threonine kinase receptor that binds BMP ligands and forms heteromeric complexes with type I receptors to initiate canonical SMAD1/5/8 signaling as well as non-canonical MAPK and cytoskeletal pathways. Through these cascades, BMPR-II regulates cell fate decisions including proliferation, differentiation, migration, and apoptosis, with prominent roles in vascular homeostasis and developmental patterning. Altered BMPR2 expression or signaling disrupts BMP/TGF-β network balance and is frequently studied in contexts of endothelial dysfunction, aberrant smooth muscle remodeling, and congenital signaling defects. BMPR-II is therefore a key node for mechanistic studies of BMP-driven transcriptional programs and pathway cross-talk in human cells.

BMPR-II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BMPR2 expression without altering the underlying DNA sequence.

BMPR-II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BMPR2 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 BMPR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BMPR-II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BMPR2 locus and enabling the study of BMPR-II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BMPR-II pathway restoration in tumor cells with silenced or reduced BMPR2 expression.

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