FLAD1 Double Nickase Plasmid (h): sc-406725-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- FLAD1 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
- FLAD1 Double Nickase Plasmid (h) and FLAD1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting FLAD1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: FLAD1 Antibody (G-4): sc-376819
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
FLAD1 Double Nickase Plasmid (h) | sc-406725-NIC | 20 µg | $410.00 |
FLAD1 encodes flavin adenine dinucleotide synthetase 1, a key enzyme that catalyzes the final step of FAD biosynthesis from FMN, thereby controlling intracellular flavin cofactor availability. FAD is required for the activity of numerous flavoproteins involved in mitochondrial oxidative metabolism, fatty acid β-oxidation, amino acid catabolism, and cellular redox homeostasis. Through its impact on electron transfer reactions and oxidative stress responses, FLAD1 is functionally linked to energy metabolism pathways and mitochondrial quality control. Altered FLAD1 function has been associated with inborn errors of metabolism featuring impaired flavoprotein-dependent processes, making it a relevant target for mechanistic studies of mitochondrial dysfunction and metabolic stress.
FLAD1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the FLAD1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within FLAD1. 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 FLAD1 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 FLAD1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.