ATP6AP1 Double Nickase Plasmid (h): sc-404162-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ATP6AP1 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
- ATP6AP1 Double Nickase Plasmid (h) and ATP6AP1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP6AP1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ATP6AP1 Antibody (E-10): sc-515607
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ATP6AP1 Double Nickase Plasmid (h) | sc-404162-NIC | 20 µg | $410.00 | |||
ATP6AP1 Double Nickase Plasmid (h2) | sc-404162-NIC-2 | 20 µg | $410.00 |
ATP6AP1 encodes an accessory component of the vacuolar H+-ATPase (V-ATPase) machinery that supports organelle acidification across the endolysosomal system. By modulating lumenal pH in compartments such as lysosomes and the Golgi, ATP6AP1 influences protein trafficking, receptor recycling, and lysosome-dependent macromolecule turnover, thereby shaping cellular proteostasis and signaling. Proper V-ATPase function intersects with autophagy-lysosome pathways and glycosylation-dependent secretory processes that are critical for maintaining homeostasis under metabolic and stress conditions. Dysregulation of ATP6AP1-associated acidification and trafficking has been linked to disorders affecting cellular metabolism, membrane transport, and organ system physiology, making it a relevant target for mechanistic studies of endomembrane function.
ATP6AP1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP6AP1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP6AP1. 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 ATP6AP1 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 ATP6AP1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.