patched/PTCH1 Double Nickase Plasmid (m): sc-422471-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- patched/PTCH1 Double Nickase Plasmid (m) 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 (m) and patched/PTCH1 Double Nickase Plasmid (m2) 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 (3B3): sc-293416
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
patched/PTCH1 Double Nickase Plasmid (m) | sc-422471-NIC | 20 µg | $410.00 | |||
patched/PTCH1 Double Nickase Plasmid (m2) | sc-422471-NIC-2 | 20 µg | $410.00 |
Ptch1 encodes the mouse patched (PTCH1) receptor, a 12-pass transmembrane protein that functions as the primary inhibitor of Smoothened in the Hedgehog signaling pathway. Binding of Hedgehog ligands such as SHH relieves PTCH1-mediated repression, enabling downstream GLI transcriptional programs that regulate embryonic patterning, stem and progenitor cell behavior, and tissue homeostasis. Disruption of PTCH1-dependent signaling control is linked to aberrant Hedgehog pathway activation and is widely studied in developmental disorders and tumor biology. Ptch1 is therefore a key node for investigating cilia-associated signal transduction, morphogen gradients, and transcriptional regulation during development and disease modeling in mouse systems.
patched/PTCH1 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Ptch1 locus in mouse 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.