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.
vinculin Double Nickase Plasmid (h): sc-400227-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- vinculin 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
- vinculin Double Nickase Plasmid (h) and vinculin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting VCL. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: vinculin Antibody (7F9): sc-73614
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
vinculin Double Nickase Plasmid (h) | sc-400227-NIC | 20 µg | $410.00 | |||
vinculin Double Nickase Plasmid (h2) | sc-400227-NIC-2 | 20 µg | $410.00 |
VCL encodes vinculin, a cytoskeletal adaptor that couples integrins and cadherins to the actin network at focal adhesions and adherens junctions, stabilizing cell–matrix and cell–cell contacts. Vinculin participates in mechanotransduction by responding to tensile force and coordinating assembly of adhesion complexes, influencing cell spreading, traction, and migration. Through interactions with talin, paxillin, α-actinin, and F-actin, it integrates signaling pathways linked to Rho family GTPases, actin remodeling, and adhesion turnover. Altered VCL expression or adhesion dynamics has been associated with impaired tissue integrity and remodeling phenotypes, and is frequently studied in contexts such as cancer cell invasion, cardiomyocyte structure, and fibrosis-related extracellular matrix responses.
vinculin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the VCL locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within VCL. 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 VCL 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 VCL-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.