GI24 Double Nickase Plasmid (h): sc-407831-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GI24 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
- GI24 Double Nickase Plasmid (h) and GI24 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting VSIR. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GI24 Double Nickase Plasmid (h) | sc-407831-NIC | 20 µg | $410.00 | |||
GI24 Double Nickase Plasmid (h2) | sc-407831-NIC-2 | 20 µg | $410.00 |
VSIR (V-set immunoregulatory receptor), also known as GI24/VISTA, encodes a B7 family immune checkpoint molecule that modulates T cell activation and helps maintain peripheral tolerance. GI24 is expressed primarily in hematopoietic and myeloid compartments, where it influences antigen-presenting cell function and dampens inflammatory cytokine programs. Through regulation of co-stimulatory signaling, VSIR contributes to immune homeostasis at tissue sites and within the tumor microenvironment. Dysregulated VSIR expression or signaling has been associated with altered anti-tumor immunity and chronic inflammatory states, making it a useful target for mechanistic studies of immunoregulation.
GI24 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the VSIR locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within VSIR. 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 VSIR 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 VSIR-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.