ATP6E Double Nickase Plasmid (h): sc-404046-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ATP6E 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
- ATP6E Double Nickase Plasmid (h) and ATP6E Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ATP6V1E1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ATP6E Antibody (G-3): sc-514143
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ATP6E Double Nickase Plasmid (h) | sc-404046-NIC | 20 µg | $410.00 | |||
ATP6E Double Nickase Plasmid (h2) | sc-404046-NIC-2 | 20 µg | $410.00 |
ATP6V1E1 encodes the E1 subunit (ATP6E) of the vacuolar H\+-ATPase (V-ATPase) V1 peripheral domain, a multi-subunit rotary ATPase that powers proton translocation to acidify endosomes, lysosomes, and other intracellular compartments. By controlling organelle pH, V-ATPase activity regulates receptor-mediated endocytosis, lysosomal degradation, autophagy, antigen processing, and vesicular trafficking, with downstream effects on nutrient sensing and signaling pathways coupled to membrane transport. Perturbation of V-ATPase subunit composition or function can disrupt cellular homeostasis and has been linked to inherited disorders of acidification and neurodevelopmental phenotypes, as well as altered proteostasis and metabolic stress responses. ATP6V1E1 is therefore a useful locus for studying pH-dependent regulation of membrane dynamics and organelle function in human cells.
ATP6E Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ATP6V1E1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ATP6V1E1. 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 ATP6V1E1 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 ATP6V1E1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.