Hhip CRISPR Activation Plasmid (h): sc-403293-ACT

Human HHIP encodes hedgehog-interacting protein (Hhip), a membrane-associated glycoprotein that binds Hedgehog ligands and attenuates pathway signaling by limiting ligand availability to PTCH1/SMO. By acting as a negative regulator of Hedgehog activity, HHIP helps tune developmental patterning programs and adult tissue homeostasis, influencing processes such as epithelial differentiation, stromal-epithelial signaling, and cellular proliferation. Dysregulated HHIP expression or altered Hedgehog pathway balance has been linked to disease-relevant phenotypes in oncology and chronic lung biology, including effects on airway remodeling and tumor-associated signaling networks. As a pathway modulator, HHIP is frequently studied in contexts where Hedgehog-driven transcriptional programs intersect with WNT, TGF-β, and inflammatory signaling.

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

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

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