INSIG-1 Double Nickase Plasmid (h): sc-401381-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- INSIG-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
- INSIG-1 Double Nickase Plasmid (h) and INSIG-1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting INSIG1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: INSIG-1 Antibody (A-9): sc-390504
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
INSIG-1 Double Nickase Plasmid (h) | sc-401381-NIC | 20 µg | $410.00 | |||
INSIG-1 Double Nickase Plasmid (h2) | sc-401381-NIC-2 | 20 µg | $410.00 |
INSIG1 encodes insulin induced gene 1 (INSIG-1), an endoplasmic reticulum membrane protein that functions as a sterol sensor and key negative regulator of lipid homeostasis. INSIG-1 binds SCAP and sterol regulatory element binding proteins (SREBPs) to retain the SCAP–SREBP complex in the ER under sterol-replete conditions, thereby limiting SREBP processing and downstream transcription of cholesterol and fatty acid biosynthetic genes. Through this control point, INSIG-1 integrates feedback regulation within the mevalonate and lipogenesis pathways and intersects with ER-associated degradation of sterol pathway components. Dysregulation of INSIG1-linked sterol sensing has been studied in metabolic phenotypes such as dyslipidemia, hepatic steatosis, and insulin resistance, and is also relevant to contexts where altered lipid metabolism supports cellular stress adaptation.
INSIG-1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the INSIG1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within INSIG1. 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 INSIG1 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 INSIG1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.