Ankyrin G Double Nickase Plasmid (h): sc-400421-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ankyrin G 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
- Ankyrin G Double Nickase Plasmid (h) and Ankyrin G Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ANK3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ankyrin G Antibody (463): sc-12719
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ankyrin G Double Nickase Plasmid (h) | sc-400421-NIC | 20 µg | $410.00 | |||
Ankyrin G Double Nickase Plasmid (h2) | sc-400421-NIC-2 | 20 µg | $410.00 |
ANK3 encodes ankyrin G, a membrane adaptor protein that links diverse ion channels and cell adhesion molecules to the spectrin–actin cytoskeleton, organizing specialized membrane domains such as the axon initial segment and nodes of Ranvier. Through regulation of channel clustering and scaffolding at excitable membranes, ankyrin G influences neuronal polarity, action potential initiation, and synaptic stability, and it also contributes to epithelial cell–cell junction integrity. ANK3-associated perturbations have been connected to neuropsychiatric disease genetics and altered neuronal network function, supporting its use as a mechanistic node in studies of excitability, circuit development, and cytoskeletal organization. These properties make ANK3 a relevant target for dissecting pathways governing membrane protein trafficking, cytoskeletal anchoring, and cell signaling at specialized subcellular compartments.
Ankyrin G Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ANK3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ANK3. 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 ANK3 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 ANK3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.