Inhibin β-A CRISPR Activation Plasmid (h): sc-402676-ACT

INHBA encodes the human Inhibin beta-A subunit, which homodimerizes to form activin A or heterodimerizes with inhibin alpha to form inhibin A, key modulators of reproductive endocrinology and tissue homeostasis. Activin/inhibin signaling engages TGF-β superfamily pathways, prominently SMAD2/3-mediated transcriptional programs that regulate cell proliferation, differentiation, extracellular matrix remodeling, and inflammatory responses. INHBA activity has been implicated in developmental processes and wound repair, and altered expression is reported across multiple disease contexts, including fibrosis and tumor-associated microenvironment remodeling. As a secreted ligand component, Inhibin beta-A also serves as a node linking paracrine signaling to downstream gene expression changes in diverse cell types.

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

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

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