Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
patched/PTCH1 Double Nickase Plasmid (h): sc-400457-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- patched/PTCH1 Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- patched/PTCH1 Double Nickase Plasmid (h) and patched/PTCH1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting PTCH1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: patched/PTCH1 Antibody (A-2): sc-518044
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
patched/PTCH1 Double Nickase Plasmid (h) | sc-400457-NIC | 20 µg | $410.00 | |||
patched/PTCH1 Double Nickase Plasmid (h2) | sc-400457-NIC-2 | 20 µg | $410.00 |
PTCH1 encodes the human patched receptor, a 12-pass transmembrane protein that serves as the primary ligand-binding and inhibitory component of the Hedgehog signaling pathway. In the absence of Hedgehog ligands, PTCH1 suppresses SMO activity to restrain GLI-dependent transcription programs that control embryonic patterning, stem/progenitor cell behavior, and tissue homeostasis. PTCH1 activity is closely linked to ciliary trafficking and signal transduction at the primary cilium, integrating developmental cues with cell-cycle and differentiation states. Genetic and regulatory perturbations of PTCH1 are associated with dysregulated Hedgehog pathway output and are widely studied in developmental disorders and Hedgehog-driven tumor biology.
patched/PTCH1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the PTCH1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within PTCH1. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt PTCH1 function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of PTCH1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.