IRS-1 Double Nickase Plasmid (h): sc-400096-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- IRS-1 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
- IRS-1 Double Nickase Plasmid (h) and IRS-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting IRS1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: IRS-1 Antibody (H-7): sc-515017
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
IRS-1 Double Nickase Plasmid (h) | sc-400096-NIC | 20 µg | $410.00 | |||
IRS-1 Double Nickase Plasmid (h2) | sc-400096-NIC-2 | 20 µg | $410.00 |
Insulin receptor substrate 1 (IRS1) encodes IRS-1, a central adaptor protein that couples activated insulin and IGF-1 receptors to downstream signaling through PI3K–AKT–mTOR and RAS–MAPK pathways. Tyrosine phosphorylation of IRS-1 coordinates recruitment of SH2-domain effectors, integrating signals that regulate glucose uptake, glycogen synthesis, protein translation, cell growth, and survival. IRS-1 function is modulated by inhibitory serine/threonine phosphorylation and feedback from mTOR/S6K, shaping insulin sensitivity and nutrient signaling dynamics. Dysregulated IRS1/IRS-1 signaling is widely studied in insulin resistance and metabolic syndrome, and altered pathway wiring is also relevant to proliferative and stress-response phenotypes in cancer and other complex diseases.
IRS-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the IRS1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within IRS1. 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 IRS1 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 IRS1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.