Villin Double Nickase Plasmid (h): sc-400587-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Villin 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
- Villin Double Nickase Plasmid (h) and Villin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting VIL1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Villin Antibody (1D2C3): sc-58897
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Villin Double Nickase Plasmid (h) | sc-400587-NIC | 20 µg | $410.00 | |||
Villin Double Nickase Plasmid (h2) | sc-400587-NIC-2 | 20 µg | $410.00 |
VIL1 encodes villin, a Ca²⁺-regulated actin-binding protein enriched in intestinal and other epithelial cells where it bundles, caps, and severs F-actin to support microvillus assembly and brush-border architecture. Through dynamic remodeling of the apical cytoskeleton, villin contributes to epithelial polarity, barrier function, and processes linked to cell migration and wound restitution. Altered VIL1 expression is widely used as a marker of epithelial differentiation, and dysregulation of villin-associated cytoskeletal programs has been studied in contexts of epithelial injury, inflammation, and tumor biology. As a scaffold for actin dynamics, villin connects calcium signaling and phosphoinositide-dependent regulation to structural maintenance of the apical surface.
Villin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the VIL1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within VIL1. 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 VIL1 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 VIL1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.