HNF-3α CRISPR Activation Plasmid (h): sc-400743-ACT

FOXA1 (HNF-3α) is a pioneer forkhead transcription factor that binds compacted chromatin to establish accessible regulatory landscapes and coordinate lineage-specific gene expression programs. In human epithelial tissues, it integrates hormone receptor signaling with transcriptional networks controlling differentiation, metabolism, and cell-cycle progression, including extensive crosstalk with nuclear receptor pathways such as estrogen and androgen receptor–dependent transcription. FOXA1 activity influences enhancer selection and chromatin remodeling, shaping developmental and homeostatic processes in liver, lung, pancreas, and mammary epithelium. Dysregulated FOXA1 expression or altered chromatin binding profiles are frequently studied in hormone-driven cancers and other diseases characterized by perturbed epithelial identity and transcriptional reprogramming.

HNF-3α CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FOXA1 expression without altering the underlying DNA sequence.

HNF-3α CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FOXA1 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 FOXA1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous HNF-3α expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FOXA1 locus and enabling the study of HNF-3α-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of HNF-3α pathway restoration in tumor cells with silenced or reduced FOXA1 expression.

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