Ep-CAM Double Nickase Plasmid (h): sc-400220-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ep-CAM 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
- Ep-CAM Double Nickase Plasmid (h) and Ep-CAM Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EPCAM. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ep-CAM Antibody (C-10): sc-25308
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ep-CAM Double Nickase Plasmid (h) | sc-400220-NIC | 20 µg | $410.00 | |||
Ep-CAM Double Nickase Plasmid (h2) | sc-400220-NIC-2 | 20 µg | $410.00 |
EPCAM encodes epithelial cell adhesion molecule (Ep-CAM), a transmembrane glycoprotein enriched at epithelial junctions that supports homotypic cell–cell adhesion and maintenance of epithelial architecture. Beyond adhesion, Ep-CAM participates in regulation of proliferation and differentiation through signaling programs that intersect with Wnt/β-catenin activity and epithelial-to-mesenchymal transition-associated processes. Altered EPCAM expression and genetic variation are linked to epithelial tumor biology, influencing cell adhesion, migration, and tissue organization in multiple carcinoma contexts. EPCAM is also widely used as a molecular marker for epithelial lineage and circulating tumor cell research, enabling studies of epithelial identity and plasticity.
Ep-CAM Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EPCAM locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EPCAM. 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 EPCAM 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 EPCAM-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.