Epac Double Nickase Plasmid (h): sc-401807-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Epac 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
- Epac Double Nickase Plasmid (h) and Epac Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RAPGEF3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Epac Antibody (A-5): sc-28366
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Epac Double Nickase Plasmid (h) | sc-401807-NIC | 20 µg | $410.00 | |||
Epac Double Nickase Plasmid (h2) | sc-401807-NIC-2 | 20 µg | $410.00 |
Human RAPGEF3 encodes Epac (exchange protein directly activated by cAMP), a guanine nucleotide exchange factor that preferentially activates Rap1 and Rap2 to couple cAMP signals to small GTPase–dependent responses. Epac regulates integrin-mediated adhesion, endothelial barrier function, vesicle trafficking, and cell junction remodeling, integrating inputs from GPCR signaling into cytoskeletal and membrane dynamics. Through Rap-driven control of MAPK and PI3K-related signaling nodes, RAPGEF3 influences proliferation, migration, and secretion across multiple cell types. Dysregulated cAMP–Epac–Rap signaling has been implicated in cardiovascular and metabolic phenotypes and is studied in contexts including inflammation, fibrosis, and cancer-associated changes in adhesion and invasiveness.
Epac Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RAPGEF3 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RAPGEF3. 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 RAPGEF3 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 RAPGEF3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.