TGIF CRISPR Activation Plasmid (h): sc-401472-ACT

Human TGIF1 encodes TGIF, a TALE homeobox transcriptional regulator that modulates gene expression programs controlling cell fate decisions during development and tissue homeostasis. TGIF functions prominently as a corepressor in TGF-β/SMAD-dependent transcription, shaping signaling outputs that influence epithelial–mesenchymal dynamics, proliferation, and differentiation. Through interactions with SMADs and additional transcriptional cofactors, TGIF helps tune pathway amplitude and duration, linking extracellular cues to chromatin-level control of target genes. Dysregulated TGIF1 activity or expression has been associated with altered TGF-β signaling states implicated in developmental abnormalities and disease-relevant transcriptional reprogramming in cancer and fibrosis research contexts.

TGIF CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TGIF1 expression without altering the underlying DNA sequence.

TGIF CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TGIF1 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 TGIF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TGIF expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TGIF1 locus and enabling the study of TGIF-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TGIF pathway restoration in tumor cells with silenced or reduced TGIF1 expression.

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