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.
SMO/Smoothened Double Nickase Plasmid (m): sc-435662-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- SMO/Smoothened 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
- SMO/Smoothened Double Nickase Plasmid (m) and SMO/Smoothened Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Smo. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: SMO/Smoothened Antibody (E-5): sc-166685
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
SMO/Smoothened Double Nickase Plasmid (m) | sc-435662-NIC | 20 µg | $410.00 | |||
SMO/Smoothened Double Nickase Plasmid (m2) | sc-435662-NIC-2 | 20 µg | $410.00 |
Mouse Smo encodes Smoothened (SMO), a seven-transmembrane signal transducer that relays Hedgehog pathway activity downstream of PTCH1 to regulate GLI-dependent transcriptional programs controlling embryonic patterning, stem and progenitor cell behavior, and tissue homeostasis. SMO activation influences primary cilium–dependent signaling events and integrates with pathways governing proliferation, differentiation, and cell fate decisions. Dysregulated Hedgehog–SMO signaling is implicated in developmental abnormalities and oncogenic processes, including aberrant growth and lineage specification in multiple tissues, making Smo a widely used node for mechanistic studies of Hedgehog pathway regulation.
SMO/Smoothened Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Smo locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Smo. 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 Smo 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 Smo-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.