Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Fe65 Double Nickase Plasmid (h): sc-403226-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Fe65 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
- Fe65 Double Nickase Plasmid (h) and Fe65 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting APBB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Fe65 Antibody (F-6): sc-398389
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Fe65 Double Nickase Plasmid (h) | sc-403226-NIC | 20 µg | $410.00 | |||
Fe65 Double Nickase Plasmid (h2) | sc-403226-NIC-2 | 20 µg | $410.00 |
APBB1 encodes Fe65, an adaptor protein that binds the amyloid precursor protein (APP) cytoplasmic tail via its phosphotyrosine-binding domains and coordinates multiprotein complexes involved in neuronal signaling. Fe65 participates in endocytic trafficking, synaptic function, and regulation of gene expression through interactions with APP processing machinery and nuclear co-regulators. These activities connect APBB1 to pathways governing APP cleavage, intracellular signaling, and activity-dependent transcription. Altered Fe65–APP interactions and downstream signaling have been studied in the context of neurodegeneration and amyloidogenic processing, supporting its relevance in mechanistic models of Alzheimer’s disease and related neuropathology.
Fe65 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the APBB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within APBB1. 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 APBB1 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 APBB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.