BMP-6 CRISPR Activation Plasmid (h): sc-401979-ACT

Human BMP6 encodes bone morphogenetic protein 6 (BMP-6), a secreted TGF-β superfamily ligand that signals through BMP type I/II receptors to activate SMAD1/5/8 and transcriptional programs controlling osteogenic differentiation, extracellular matrix remodeling, and developmental patterning. BMP-6 also interfaces with MAPK and PI3K-related signaling to influence cell fate decisions and tissue homeostasis in a context-dependent manner. In the liver, BMP-6 is a key upstream regulator of iron metabolism via induction of hepcidin (HAMP), linking BMP/SMAD signaling to systemic iron balance. Dysregulated BMP6 pathway activity has been associated with altered bone biology, fibrosis-related processes, and iron homeostasis phenotypes, making it relevant for mechanistic studies of BMP signaling and metabolic regulation.

BMP-6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous BMP6 expression without altering the underlying DNA sequence.

BMP-6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the BMP6 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 BMP6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous BMP-6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native BMP6 locus and enabling the study of BMP-6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of BMP-6 pathway restoration in tumor cells with silenced or reduced BMP6 expression.

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