Ebi2 Double Nickase Plasmid (h): sc-405397-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ebi2 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
- Ebi2 Double Nickase Plasmid (h) and Ebi2 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting GPR183. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Ebi2 Antibody (G-12): sc-514342
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ebi2 Double Nickase Plasmid (h) | sc-405397-NIC | 20 µg | $410.00 | |||
Ebi2 Double Nickase Plasmid (h2) | sc-405397-NIC-2 | 20 µg | $410.00 |
GPR183 encodes EBI2 (also known as GPR183), a G protein–coupled receptor that senses oxysterols such as 7α,25-dihydroxycholesterol to guide immune cell positioning. EBI2 signaling coordinates chemotactic responses and contributes to lymphoid tissue organization by shaping B cell and dendritic cell localization within follicles and interfollicular regions. Through GPCR-mediated pathways including Gαi-dependent signaling, it influences migration, activation states, and antigen-driven interactions that are central to humoral immunity. Dysregulated GPR183 activity has been implicated in inflammatory and autoimmune contexts and has been explored in infection-related immune remodeling, making it a useful node for studying immune cell trafficking and oxysterol biology.
Ebi2 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the GPR183 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within GPR183. 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 GPR183 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 GPR183-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.