FAT1 Double Nickase Plasmid (h): sc-410778-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FAT1 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
- FAT1 Double Nickase Plasmid (h) and FAT1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FAT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FAT1 Antibody (1634CT464.1.9): sc-517329
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FAT1 Double Nickase Plasmid (h) | sc-410778-NIC | 20 µg | $410.00 | |||
FAT1 Double Nickase Plasmid (h2) | sc-410778-NIC-2 | 20 µg | $410.00 |
FAT1 encodes an atypical cadherin that functions as a large transmembrane cell-adhesion and polarity regulator, coordinating cytoskeletal organization, planar cell polarity–like processes, and contact-dependent signaling. FAT1 has been linked to modulation of Hippo pathway output and crosstalk with Wnt/β-catenin–associated programs, influencing cell migration, tissue architecture, and mechanotransduction. Dysregulated FAT1 activity or mutation has been reported across multiple tumor types and is also associated with developmental and renal phenotypes, supporting its broad relevance to studies of morphogenesis and epithelial integrity. In cultured human cells, FAT1 perturbation is commonly used to interrogate adhesion-dependent signaling, invasion dynamics, and polarity control.
FAT1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FAT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FAT1. 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 FAT1 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 FAT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.