AP-2γ CRISPR Activation Plasmid (h): sc-400884-ACT

TFAP2C encodes the human transcription factor AP-2γ (TFAP2C), a sequence-specific DNA-binding protein that regulates gene expression programs controlling epithelial differentiation, proliferation, and lineage commitment. AP-2γ integrates signals across developmental and hormone-responsive pathways and modulates transcriptional networks involved in cell cycle progression, apoptosis, and chromatin-state regulation. Dysregulated TFAP2C activity has been associated with altered epithelial identity, invasiveness, and transcriptional reprogramming in multiple cancer contexts, making it a useful node for studying oncogenic transcriptional circuitry. In addition, TFAP2C contributes to placental and mammary epithelial biology, providing a framework for interrogating tissue-specific enhancer usage and transcription factor cooperativity.

AP-2γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TFAP2C expression without altering the underlying DNA sequence.

AP-2γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TFAP2C 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 TFAP2C transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous AP-2γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TFAP2C locus and enabling the study of AP-2γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of AP-2γ pathway restoration in tumor cells with silenced or reduced TFAP2C expression.

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