Gα i-3 Double Nickase Plasmid (h): sc-400453-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Gα i-3 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 i-3 Double Nickase Plasmid (h) and Gα i-3 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GNAI3. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Gα i-3 Double Nickase Plasmid (h) | sc-400453-NIC | 20 µg | $410.00 | |||
Gα i-3 Double Nickase Plasmid (h2) | sc-400453-NIC-2 | 20 µg | $410.00 |
GNAI3 encodes the human heterotrimeric G protein alpha subunit Gαi-3, a GDP/GTP-regulated molecular switch that couples many GPCRs to inhibition of adenylyl cyclase and reduced cAMP signaling. Through downstream effects on PKA activity, ion channels, and MAPK/ERK and PI3K-associated networks, Gαi-3 helps coordinate chemotaxis, secretion, and cell growth decisions in response to extracellular cues. It also participates in membrane trafficking and polarity programs by regulating vesicle transport and cytoskeletal dynamics. Altered GPCR–Gαi signaling has been associated with dysregulated proliferative and migratory behavior in disease-relevant cellular contexts, supporting mechanistic studies of pathway rewiring.
Gα i-3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GNAI3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GNAI3. 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 GNAI3 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 GNAI3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.