TGFβ2 CRISPR Activation Plasmid (h): sc-400496-ACT

TGFB2 encodes transforming growth factor beta 2 (TGFβ2), a secreted cytokine that signals through TGF-β type I/II serine-threonine kinase receptors to activate SMAD-dependent transcription and non-canonical pathways including MAPK, PI3K/AKT, and Rho GTPase signaling. TGFβ2 regulates extracellular matrix deposition, epithelial–mesenchymal transition, cell fate decisions, immune modulation, and tissue morphogenesis, contributing to homeostasis and wound repair. Dysregulated TGFB2 signaling has been implicated in fibrosis and aberrant stromal remodeling, as well as context-dependent roles in tumor progression, invasion, and immune evasion. These functions make TGFB2 a key node for dissecting microenvironmental signaling, differentiation programs, and pathway crosstalk in human cell models.

TGFβ2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TGFB2 expression without altering the underlying DNA sequence.

TGFβ2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TGFB2 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 TGFB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TGFβ2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TGFB2 locus and enabling the study of TGFβ2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TGFβ2 pathway restoration in tumor cells with silenced or reduced TGFB2 expression.

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