TGF beta 1 CRISPR Activation Plasmid (m): sc-423364-ACT

Mouse Tgfb1 encodes transforming growth factor beta 1 (TGF beta 1), a secreted cytokine that signals primarily through TGFBR1/2 to activate SMAD2/3-dependent transcription and intersect with non-canonical MAPK, PI3K/AKT, and Rho GTPase pathways. TGF beta 1 is a central regulator of extracellular matrix remodeling, epithelial-to-mesenchymal transition, immune cell differentiation, and maintenance of tissue homeostasis. Dysregulated Tgfb1 activity is broadly implicated in fibrotic remodeling, aberrant immune suppression or inflammation, impaired wound repair, and tumor–stroma interactions. In mouse models, Tgfb1 perturbation is widely used to interrogate stromal signaling, macrophage and T cell polarization, and context-dependent growth control in development and disease-like phenotypes.

TGF beta 1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Tgfb1 expression without altering the underlying DNA sequence.

TGF beta 1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Tgfb1 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 Tgfb1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TGF beta 1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Tgfb1 locus and enabling the study of TGF beta 1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TGF beta 1 pathway restoration in tumor cells with silenced or reduced Tgfb1 expression.

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