EEA1 Double Nickase Plasmid (h): sc-400234-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EEA1 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
- EEA1 Double Nickase Plasmid (h) and EEA1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EEA1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EEA1 Antibody (G-4): sc-137130
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EEA1 Double Nickase Plasmid (h) | sc-400234-NIC | 20 µg | $410.00 | |||
EEA1 Double Nickase Plasmid (h2) | sc-400234-NIC-2 | 20 µg | $410.00 |
EEA1 encodes early endosome antigen 1, a Rab5 effector that functions as a tethering factor required for early endosome docking and homotypic fusion. Through its FYVE domain, EEA1 binds phosphatidylinositol 3-phosphate and coordinates endosomal membrane identity with vesicle trafficking events, influencing receptor-mediated endocytosis and cargo sorting. EEA1-dependent endosomal dynamics intersect with PI3K signaling, cytoskeletal organization, and maturation of endocytic compartments that regulate growth factor receptor turnover. Dysregulated endosome trafficking and Rab5–EEA1 axis perturbations have been linked to altered signaling homeostasis in cancer and to endolysosomal defects implicated in neurodegenerative and infectious disease research contexts.
EEA1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EEA1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EEA1. 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 EEA1 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 EEA1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.