HNF-3γ CRISPR Activation Plasmid (h): sc-401720-ACT

FOXA3 encodes hepatocyte nuclear factor 3 gamma (HNF-3γ), a forkhead box transcription factor that helps establish and maintain endodermal lineage programs and regulates genes involved in epithelial differentiation and metabolic homeostasis. HNF-3γ participates in transcriptional networks that coordinate chromatin accessibility and promoter/enhancer activity, intersecting with pathways controlling glucose and lipid metabolism, hepatic function, and gastrointestinal development. In human cells, FOXA3 activity contributes to cell identity decisions and context-dependent remodeling of gene regulatory circuits. Dysregulated FOXA3/HNF-3γ expression or downstream transcriptional programs has been associated with altered differentiation states and disease-relevant phenotypes in liver and gastrointestinal biology, making it a useful node for mechanistic studies.

HNF-3γ CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXA3 expression without altering the underlying DNA sequence.

HNF-3γ CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXA3 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 FOXA3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HNF-3γ expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXA3 locus and enabling the study of HNF-3γ-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HNF-3γ pathway restoration in tumor cells with silenced or reduced FOXA3 expression.

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