Inhibin β-B CRISPR Activation Plasmid (h): sc-402060-ACT

INHBB encodes the human Inhibin beta-B subunit, a TGF-β superfamily ligand that forms activin B homodimers or activin AB heterodimers to regulate cell fate decisions and endocrine signaling. Activins signal primarily through type II/I serine-threonine kinase receptors to activate SMAD2/3-dependent transcriptional programs, influencing proliferation, differentiation, extracellular matrix remodeling, and inflammatory responses in a context-dependent manner. INHBB activity is relevant to reproductive biology and gonadal function, and altered activin/inhibin balance has been associated with dysregulated tissue growth and remodeling in diverse pathological states, including fibrosis and cancer-related signaling networks. Because INHBB integrates paracrine and autocrine cues, it is frequently studied in pathways controlling developmental programs and microenvironment-mediated regulation of gene expression.

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

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

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