Amphiphysin II Double Nickase Plasmid (h): sc-401263-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Amphiphysin II 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
- Amphiphysin II Double Nickase Plasmid (h) and Amphiphysin II Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting BIN1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Amphiphysin II Antibody (2F11): sc-23918
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Amphiphysin II Double Nickase Plasmid (h) | sc-401263-NIC | 20 µg | $410.00 | |||
Amphiphysin II Double Nickase Plasmid (h2) | sc-401263-NIC-2 | 20 µg | $410.00 |
BIN1 encodes amphiphysin II, a BAR domain-containing membrane remodeling protein that couples phosphoinositide binding to curvature sensing and clathrin-mediated endocytosis. Amphiphysin II participates in endocytic vesicle formation, T-tubule biogenesis in muscle, and trafficking of receptors and signaling complexes, linking membrane dynamics to cytoskeletal organization. Through SH3-mediated interactions with partners such as dynamin and endocytic adaptors, BIN1 influences membrane fission and compartmentalization that shape intracellular signaling and synaptic function. Altered BIN1 expression or isoform usage has been associated with neuromuscular phenotypes and is genetically implicated in neurodegeneration, supporting its utility in modeling disease-relevant membrane trafficking defects in human cells.
Amphiphysin II Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the BIN1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within BIN1. 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 BIN1 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 BIN1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.