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 5 Double Nickase Plasmid (h): sc-400671-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Cytokeratin 5 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 5 Double Nickase Plasmid (h) and Cytokeratin 5 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KRT5. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Cytokeratin 5 Antibody (RCK103): sc-32721
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Cytokeratin 5 Double Nickase Plasmid (h) | sc-400671-NIC | 20 µg | $410.00 | |||
Cytokeratin 5 Double Nickase Plasmid (h2) | sc-400671-NIC-2 | 20 µg | $410.00 |
KRT5 encodes cytokeratin 5, a type II intermediate filament protein that heterodimerizes with keratin 14 to form the basal keratin network in stratified epithelia. This cytoskeletal scaffold supports mechanical resilience, cell polarity, and attachment structures such as hemidesmosomes, integrating with programs that regulate epithelial differentiation and barrier maintenance. KRT5 expression is widely used to mark basal keratinocytes and progenitor-like epithelial states, and altered KRT5 biology is implicated in epithelial fragility phenotypes and dysregulated differentiation in skin and airway contexts. Consequently, KRT5 is frequently studied in models of epidermal homeostasis, wound responses, and epithelial remodeling relevant to disease-associated epithelial stress and degeneration.
Cytokeratin 5 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KRT5 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KRT5. 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 KRT5 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 KRT5-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.