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.
Cingulin Double Nickase Plasmid (h): sc-404409-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cingulin 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
- Cingulin Double Nickase Plasmid (h) and Cingulin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CGN. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cingulin Antibody (G-6): sc-365264
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cingulin Double Nickase Plasmid (h) | sc-404409-NIC | 20 µg | $410.00 | |||
Cingulin Double Nickase Plasmid (h2) | sc-404409-NIC-2 | 20 µg | $410.00 |
CGN encodes cingulin, a cytoplasmic scaffolding protein concentrated at tight junctions where it links transmembrane junctional components to the cortical actin cytoskeleton. Cingulin coordinates epithelial barrier formation and maintenance by modulating junction assembly, cell polarity, and cytoskeletal organization, and it interfaces with signaling pathways that regulate junctional tension and Rho-family GTPase activity. Through these roles, CGN influences paracellular permeability and tissue homeostasis in polarized epithelia and endothelia. Altered tight junction architecture and cingulin-associated regulatory networks are frequently investigated in contexts of barrier dysfunction, inflammation, and epithelial remodeling relevant to complex disease phenotypes.
Cingulin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CGN locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CGN. 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 CGN 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 CGN-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.