ATP5F1 Double Nickase Plasmid (h): sc-405710-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ATP5F1 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
- ATP5F1 Double Nickase Plasmid (h) and ATP5F1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP5F1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ATP5F1 Antibody (C-12): sc-514419
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ATP5F1 Double Nickase Plasmid (h) | sc-405710-NIC | 20 µg | $410.00 | |||
ATP5F1 Double Nickase Plasmid (h2) | sc-405710-NIC-2 | 20 µg | $410.00 |
ATP5F1 encodes the catalytic core of mitochondrial F1F0 ATP synthase (Complex V), a key enzyme that couples oxidative phosphorylation to ATP production across the inner mitochondrial membrane. By converting the proton motive force into cellular ATP, ATP5F1 supports energy-dependent processes including ion homeostasis, biosynthesis, and stress adaptation, and it integrates with electron transport chain activity and mitochondrial membrane potential control. Perturbation of ATP synthase function is linked to mitochondrial dysfunction, altered reactive oxygen species handling, and metabolic reprogramming that can influence cell survival and proliferation. As a central node in bioenergetics, ATP5F1 is frequently studied in the context of neuromuscular and neurodegenerative phenotypes, as well as mitochondrial contributions to cancer cell metabolism.
ATP5F1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP5F1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP5F1. 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 ATP5F1 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 ATP5F1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.