Gα s Double Nickase Plasmid (h): sc-400476-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Gα s 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
- Gα s Double Nickase Plasmid (h) and Gα s Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GNAS. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Gα s Antibody (12): sc-135914
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Gα s Double Nickase Plasmid (h) | sc-400476-NIC | 20 µg | $410.00 | |||
Gα s Double Nickase Plasmid (h2) | sc-400476-NIC-2 | 20 µg | $410.00 |
GNAS encodes the stimulatory heterotrimeric G protein alpha subunit Gαs, a central mediator of GPCR signaling that activates adenylyl cyclase to elevate cAMP and stimulate PKA- and EPAC-dependent transcriptional and metabolic programs. Through coupling to diverse receptors, Gαs regulates hormone responsiveness, ion transport, and cell-state decisions, intersecting with CREB signaling and feedback control of second-messenger dynamics. Dysregulated GNAS activity and imprinting have been implicated in endocrine and metabolic phenotypes, and altered cAMP signaling is frequently examined for roles in growth control and differentiation. As a hub in GPCR–cAMP pathways, Gαs is commonly studied to map receptor coupling, pathway cross-talk, and context-dependent transcriptional responses.
Gα s Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GNAS locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GNAS. 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 GNAS 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 GNAS-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.