WT1 Double Nickase Plasmid (h): sc-400095-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- WT1 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
- WT1 Double Nickase Plasmid (h) and WT1 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting WT1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: WT1 Antibody (H-1): sc-393498
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
WT1 Double Nickase Plasmid (h) | sc-400095-NIC | 20 µg | $410.00 | |||
WT1 Double Nickase Plasmid (h2) | sc-400095-NIC-2 | 20 µg | $410.00 |
WT1 encodes a zinc-finger transcription factor that regulates lineage specification, cell-cycle control, and apoptosis through context-dependent transcriptional activation or repression. It functions in developmental programs, particularly urogenital and hematopoietic tissues, and interfaces with pathways controlling differentiation and epithelial–mesenchymal plasticity via modulation of growth factor and transcriptional networks. WT1 also participates in RNA processing through isoform-specific effects on splicing and nucleic acid binding, contributing to complex regulatory outputs. Dysregulated WT1 expression or mutation is implicated in multiple malignancies and developmental disorders, making it a widely used marker and mechanistic node in studies of oncogenic transcriptional circuitry and cell fate regulation.
WT1 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the WT1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within WT1. 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 WT1 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 WT1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.