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.
Cytokeratin 18 Double Nickase Plasmid (h): sc-400305-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cytokeratin 18 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
- Cytokeratin 18 Double Nickase Plasmid (h) and Cytokeratin 18 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KRT18. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cytokeratin 18 Antibody (RGE53): sc-32329
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cytokeratin 18 Double Nickase Plasmid (h) | sc-400305-NIC | 20 µg | $410.00 | |||
Cytokeratin 18 Double Nickase Plasmid (h2) | sc-400305-NIC-2 | 20 µg | $410.00 |
KRT18 encodes cytokeratin 18, a type I intermediate filament protein that heteropolymerizes with keratin 8 to form the core cytoskeletal network of simple epithelial cells. Cytokeratin 18 supports cellular mechanical stability, organelle positioning, and epithelial polarity, and it is dynamically remodeled during mitosis and stress responses via phosphorylation-dependent filament reorganization. It participates in apoptosis-associated cytoskeletal changes, including caspase-mediated cleavage that can influence epithelial cell integrity and signaling. Altered KRT18 expression, filament organization, or cleavage patterns have been associated with epithelial injury and a range of pathological states, making it a common marker and mechanistic node in studies of epithelial homeostasis and stress.
Cytokeratin 18 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KRT18 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KRT18. 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 KRT18 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 KRT18-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.