ANO1 Double Nickase Plasmid (h): sc-401314-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ANO1 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
- ANO1 Double Nickase Plasmid (h) and ANO1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ANO1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: ANO1 Antibody (C-5): sc-377115
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ANO1 Double Nickase Plasmid (h) | sc-401314-NIC | 20 µg | $410.00 | |||
ANO1 Double Nickase Plasmid (h2) | sc-401314-NIC-2 | 20 µg | $410.00 |
ANO1 (also known as TMEM16A) encodes a calcium-activated chloride channel that regulates epithelial fluid secretion, membrane excitability, and smooth muscle tone by coupling intracellular Ca²⁺ signals to anion flux. ANO1 activity shapes transepithelial transport, contributes to sensory and neuronal signaling, and intersects with calcium-dependent pathways that influence cell volume regulation and membrane potential. Altered ANO1 expression or function has been linked to disorders of epithelial secretion and airway physiology, and it is frequently studied in the context of tumor biology where ion channel remodeling can affect proliferation, migration, and the tumor microenvironment. These features make ANO1 a useful target for dissecting chloride channel–dependent signaling and calcium-driven physiological processes in human cells.
ANO1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ANO1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ANO1. 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 ANO1 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 ANO1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.