TGFβ3 CRISPR Activation Plasmid (h): sc-400768-ACT

TGFB3 encodes transforming growth factor beta 3 (TGFβ3), a secreted cytokine of the TGF-β superfamily that signals primarily through TGFBR1/2 to activate SMAD2/3 and modulate transcriptional programs controlling proliferation, differentiation, extracellular matrix remodeling, and immune regulation. TGFβ3 also intersects with non-canonical pathways including MAPK, PI3K/AKT, and Rho-like GTPase signaling, influencing epithelial–mesenchymal transition, wound repair, and tissue homeostasis. Dysregulated TGFB3/TGF-β signaling is implicated in fibrosis-associated remodeling, aberrant developmental patterning, and tumor microenvironment dynamics, where altered signaling can shape stromal responses and immune suppression. These functions make TGFB3 a key node for studying context-dependent regulation of cell fate and matrix biology in human systems.

TGFβ3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TGFB3 expression without altering the underlying DNA sequence.

TGFβ3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TGFB3 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 TGFB3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TGFβ3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TGFB3 locus and enabling the study of TGFβ3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TGFβ3 pathway restoration in tumor cells with silenced or reduced TGFB3 expression.

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