Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
B7-2 Double Nickase Plasmid (h): sc-418032-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- B7-2 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
- B7-2 Double Nickase Plasmid (h) and B7-2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting CD86. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: B7-2 Antibody (D-6): sc-28347
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
B7-2 Double Nickase Plasmid (h) | sc-418032-NIC | 20 µg | $410.00 | |||
B7-2 Double Nickase Plasmid (h2) | sc-418032-NIC-2 | 20 µg | $410.00 |
CD86 (B7-2) is a co-stimulatory immunoregulatory ligand expressed primarily on antigen-presenting cells that binds CD28 and CTLA-4 to tune T cell activation, anergy, and immune checkpoint balance. Through integration with TCR signaling, CD86 contributes to immunological synapse formation and downstream pathways including NF-κB, MAPK, and PI3K/AKT, shaping cytokine production and adaptive immune priming. Altered CD86 expression has been reported across inflammatory and autoimmune contexts and in tumor-immune microenvironments, where it can influence antigen presentation and the amplitude of lymphocyte responses. As a result, CD86 is frequently studied in assays of dendritic cell maturation, macrophage polarization, and T cell co-stimulation dynamics.
B7-2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the CD86 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within CD86. 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 CD86 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 CD86-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.