EGFR Double Nickase Plasmid (h): sc-400015-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EGFR 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
- EGFR Double Nickase Plasmid (h) and EGFR Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting EGFR. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EGFR Antibody (A-10): sc-373746
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EGFR Double Nickase Plasmid (h) | sc-400015-NIC | 20 µg | $410.00 | |||
EGFR Double Nickase Plasmid (h2) | sc-400015-NIC-2 | 20 µg | $410.00 |
Epidermal growth factor receptor (EGFR) is a receptor tyrosine kinase that regulates cell proliferation, survival, differentiation, and migration through ligand-induced dimerization and autophosphorylation. Activated EGFR propagates signaling through major pathways including RAS–RAF–MEK–ERK (MAPK), PI3K–AKT–mTOR, PLCγ–PKC, and JAK–STAT, integrating extracellular cues with transcriptional and metabolic programs. In human biology, dysregulated EGFR signaling via overexpression, amplification, or activating mutations is frequently associated with oncogenic phenotypes and altered responsiveness to growth factor stimuli. EGFR also participates in epithelial tissue homeostasis and wound repair, making it a central node for studying receptor signaling dynamics, trafficking, and pathway crosstalk.
EGFR Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the EGFR locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within EGFR. 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 EGFR 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 EGFR-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.