TFII-I CRISPR Activation Plasmid (h): sc-403153-ACT

GTF2I encodes TFII-I, a multifunctional transcription factor that binds initiator and E-box–like elements to integrate receptor-proximal signals with RNA polymerase II–dependent gene expression programs. TFII-I participates in growth factor and immune signaling networks, modulating pathways linked to MAPK/ERK activity, calcium-dependent transcription, and stimulus-responsive chromatin regulation. Through its roles in controlling proliferation, differentiation, and stress responses, TFII-I has been studied in the context of neurodevelopmental phenotypes and immune dysregulation, including dosage-sensitive genomic alterations at 7q11.23. Altered GTF2I regulation also provides a mechanistic entry point for investigating transcriptional rewiring in cancer-associated signaling and cell-state transitions.

TFII-I CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GTF2I expression without altering the underlying DNA sequence.

TFII-I CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GTF2I 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 GTF2I transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous TFII-I expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GTF2I locus and enabling the study of TFII-I-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of TFII-I pathway restoration in tumor cells with silenced or reduced GTF2I expression.

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