Keratin 2 Double Nickase Plasmid (h): sc-402077-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Keratin 2 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
- Keratin 2 Double Nickase Plasmid (h) and Keratin 2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting KRT2. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Keratin 2 Double Nickase Plasmid (h) | sc-402077-NIC | 20 µg | $410.00 | |||
Keratin 2 Double Nickase Plasmid (h2) | sc-402077-NIC-2 | 20 µg | $410.00 |
KRT2 encodes keratin 2, a type II intermediate filament protein predominantly expressed in suprabasal keratinocytes where it heterodimerizes with type I keratins to form the cytoskeletal network of the epidermis. Keratin 2 contributes to mechanical resilience, cell–cell cohesion, and proper differentiation during epidermal stratification, integrating with desmosome-associated junctional complexes and stress-response remodeling of intermediate filaments. Altered KRT2 function disrupts filament organization and epidermal integrity, and pathogenic variants are linked to keratinization disorders including superficial epidermolytic ichthyosis. Because keratin filament dynamics intersect with pathways controlling barrier formation, cornification, and responses to mechanical or inflammatory stress, KRT2 is a useful target for studying skin homeostasis and cytoskeletal regulation.
Keratin 2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the KRT2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within KRT2. 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 KRT2 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 KRT2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.