Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Squalene synthetase Double Nickase Plasmid (h): sc-403845-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Squalene synthetase 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
- Squalene synthetase Double Nickase Plasmid (h) and Squalene synthetase Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FDFT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Squalene synthetase Antibody (A-7): sc-271602
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Squalene synthetase Double Nickase Plasmid (h) | sc-403845-NIC | 20 µg | $410.00 | |||
Squalene synthetase Double Nickase Plasmid (h2) | sc-403845-NIC-2 | 20 µg | $410.00 |
FDFT1 encodes squalene synthetase, an endoplasmic reticulum–associated enzyme that catalyzes the first committed step in sterol biosynthesis by converting two molecules of farnesyl diphosphate into squalene. This reaction links the mevalonate pathway to downstream cholesterol production, coordinating membrane biogenesis, lipid raft organization, and synthesis of steroid-derived metabolites. By controlling flux into sterol synthesis, squalene synthetase influences cellular lipid homeostasis and feedback regulation of cholesterol-responsive transcriptional programs. Dysregulation of FDFT1-dependent sterol metabolism has been investigated in contexts of metabolic and cardiovascular biology, neurobiology, and proliferative states where mevalonate pathway activity is altered.
Squalene synthetase Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FDFT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FDFT1. 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 FDFT1 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 FDFT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.