Hex CRISPR Activation Plasmid (h): sc-403827-ACT

Human HHEX encodes Hex, a homeobox transcription factor that regulates lineage specification and organogenesis, with prominent roles in early endoderm patterning, hematopoietic development, and vascular biology. Hex coordinates transcriptional programs controlling cell fate decisions, proliferation, and differentiation through sequence-specific DNA binding and interaction with coregulatory complexes. In adult tissues, HHEX contributes to maintenance of immune and metabolic homeostasis, and dysregulated expression has been linked to altered hematopoietic differentiation and oncogenic transcriptional states. Genetic and expression studies also associate HHEX with cardiometabolic traits, supporting its use as a mechanistic node in endocrine and inflammatory pathway research.

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

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

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