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.
Riboflavin kinase Double Nickase Plasmid (m): sc-424854-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Riboflavin kinase Double Nickase Plasmid (m) 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
- Riboflavin kinase Double Nickase Plasmid (m) and Riboflavin kinase Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Rfk. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Riboflavin kinase Antibody (E-7): sc-398830
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Riboflavin kinase Double Nickase Plasmid (m) | sc-424854-NIC | 20 µg | $410.00 |
Mouse Rfk encodes riboflavin kinase, the cytosolic enzyme that phosphorylates riboflavin (vitamin B2) to flavin mononucleotide (FMN), a precursor for flavin adenine dinucleotide (FAD). By controlling cellular FMN/FAD pools, RFK supports flavoprotein-dependent redox reactions central to mitochondrial oxidative metabolism, fatty acid oxidation, and antioxidant defense. Altered flavin homeostasis can perturb electron transport chain function and reactive oxygen species handling, linking RFK-regulated metabolism to cellular stress responses. Rfk is therefore of interest in models of metabolic imbalance and neurodevelopmental and neuromuscular vulnerability driven by disrupted cofactor availability and mitochondrial dysfunction.
Riboflavin kinase Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Rfk locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Rfk. 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 Rfk 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 Rfk-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.