Duffy Double Nickase Plasmid (h): sc-406288-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Duffy 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
- Duffy Double Nickase Plasmid (h) and Duffy Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting ACKR1. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Duffy Double Nickase Plasmid (h) | sc-406288-NIC | 20 µg | $410.00 | |||
Duffy Double Nickase Plasmid (h2) | sc-406288-NIC-2 | 20 µg | $410.00 |
ACKR1 encodes the Duffy antigen receptor for chemokines (DARC), an atypical chemokine receptor expressed prominently on venular endothelial cells and erythrocytes. Duffy binds and sequesters multiple inflammatory chemokines from the CC and CXC families, shaping chemokine gradients that regulate leukocyte trafficking, endothelial activation, and inflammatory resolution. Through its role as a chemokine sink and transporter, Duffy modulates inflammatory signaling networks and influences vascular microenvironmental responses. Genetic variation and altered expression of ACKR1 are used to study differences in chemokine homeostasis and immune-cell recruitment in inflammatory and hematologic disease contexts.
Duffy Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the ACKR1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within ACKR1. 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 ACKR1 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 ACKR1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.