Wolframin Double Nickase Plasmid (h): sc-403869-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Wolframin 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
- Wolframin Double Nickase Plasmid (h) and Wolframin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting WFS1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Wolframin Double Nickase Plasmid (h) | sc-403869-NIC | 20 µg | $410.00 | |||
Wolframin Double Nickase Plasmid (h2) | sc-403869-NIC-2 | 20 µg | $410.00 |
WFS1 encodes wolframin, a multi-pass endoplasmic reticulum (ER) membrane protein that supports ER homeostasis and protects cells from maladaptive unfolded protein response signaling during secretory and metabolic stress. Wolframin participates in processes linked to ER Ca²⁺ handling, proteostasis, and crosstalk between ER stress pathways and mitochondrial function, influencing cellular survival and insulin secretory capacity. Dysregulation of WFS1 is associated with Wolfram syndrome and has been implicated in broader neurodegeneration and diabetes-related phenotypes, making it a useful node for studying stress-responsive signaling networks. In human cell models, WFS1 perturbation provides a readout for ER stress/UPR markers, calcium-dependent signaling, and downstream effects on metabolism and cell fate.
Wolframin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the WFS1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within WFS1. 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 WFS1 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 WFS1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.