GPR40 Double Nickase Plasmid (m): sc-433245-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GPR40 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
- GPR40 Double Nickase Plasmid (m) and GPR40 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Ffar1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GPR40 Double Nickase Plasmid (m) | sc-433245-NIC | 20 µg | $410.00 |
Mouse Ffar1 encodes GPR40, a G protein-coupled receptor activated by medium- and long-chain free fatty acids that links nutrient sensing to intracellular signaling. In pancreatic islets and other metabolic tissues, GPR40 couples primarily to Gq/11 to stimulate phospholipase C, elevate intracellular Ca²⁺, and engage MAPK pathways, shaping stimulus-secretion coupling and broader lipid-responsive transcriptional programs. This receptor participates in fatty acid–dependent regulation of glucose homeostasis and intersects with pathways implicated in metabolic stress, lipotoxicity, and inflammation. Altered FFAR1/GPR40 signaling has been studied in the context of obesity-associated insulin dysregulation and related metabolic phenotypes in mouse models.
GPR40 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Ffar1 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Ffar1. 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 Ffar1 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 Ffar1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.