
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Shh CRISPR Activation Plasmid (h) | sc-400225-ACT | 20 µg | $397.00 | |||
Shh CRISPR Activation Plasmid (h2) | sc-400225-ACT-2 | 20 µg | $397.00 |
Human SHH encodes Sonic hedgehog (Shh), a secreted morphogen that orchestrates embryonic patterning, tissue polarity, and stem/progenitor cell behavior through the Hedgehog signaling cascade. Shh binds PTCH1 to relieve repression of SMO, enabling GLI transcription factor activation and downstream programs governing proliferation, differentiation, and spatial organization. In adult tissues, SHH contributes to epithelial–mesenchymal interactions and niche maintenance, and dysregulated Hedgehog signaling is implicated in developmental disorders and oncogenic signaling contexts. SHH pathway activity is commonly studied for its roles in neural tube and limb development, organogenesis, and signal-dependent transcriptional control.
Shh CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SHH expression without altering the underlying DNA sequence.
Shh CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SHH 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 SHH transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Shh expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SHH locus and enabling the study of Shh-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Shh pathway restoration in tumor cells with silenced or reduced SHH expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.