V-ATPase α1 Double Nickase Plasmid (h): sc-403882-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- V-ATPase α1 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
- V-ATPase α1 Double Nickase Plasmid (h) and V-ATPase α1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP6V1A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: V-ATPase α1 Antibody (4F5): sc-293336
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
V-ATPase α1 Double Nickase Plasmid (h) | sc-403882-NIC | 20 µg | $410.00 | |||
V-ATPase α1 Double Nickase Plasmid (h2) | sc-403882-NIC-2 | 20 µg | $410.00 |
ATP6V1A encodes the catalytic A subunit of the V1 domain of vacuolar H+-ATPase (V-ATPase) α1, a core component of the ATP-driven proton pump that acidifies endosomes, lysosomes, and secretory vesicles. By controlling organellar pH, V-ATPase α1 supports receptor-mediated endocytosis, autophagic flux, lysosomal degradation, vesicular trafficking, and coupling to mTORC1 nutrient-sensing at the lysosomal surface. Proper V-ATPase activity is also required for Golgi and endosomal processing of proteins and lipids, influencing antigen presentation and signaling turnover. Dysregulated V-ATPase-dependent acidification has been linked to neurodegeneration, tumor cell metabolic adaptation and invasion, and defects in proteostasis and synaptic function, making ATP6V1A a useful target for pathway-focused mechanistic studies.
V-ATPase α1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP6V1A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP6V1A. 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 ATP6V1A 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 ATP6V1A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.