RBP-Jκ Double Nickase Plasmid (m): sc-422626-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RBP-Jκ Double Nickase Plasmid (m) 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
- RBP-Jκ Double Nickase Plasmid (m) and RBP-Jκ Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Rbpj. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RBP-Jκ Antibody (E-7): sc-271128
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RBP-Jκ Double Nickase Plasmid (m) | sc-422626-NIC | 20 µg | $410.00 | |||
RBP-Jκ Double Nickase Plasmid (m2) | sc-422626-NIC-2 | 20 µg | $410.00 |
Rbpj encodes RBP-Jκ, a sequence-specific DNA-binding transcription factor that serves as the central nuclear effector of canonical Notch signaling. Upon Notch receptor activation, RBP-Jκ switches from a transcriptional repressor to an activator through recruitment of NICD and coactivators, regulating genes controlling cell fate decisions, proliferation, and differentiation. In mouse systems, RBP-Jκ-dependent programs are critical in hematopoiesis, neurogenesis, and immune cell development, and dysregulation of Notch–RBP-Jκ transcriptional outputs is frequently used as a mechanistic framework for studying oncogenic transformation and developmental phenotypes. As a hub integrating Notch inputs with chromatin and transcriptional control, Rbpj is widely investigated in pathways governing stem/progenitor maintenance and lineage commitment.
RBP-Jκ Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Rbpj locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Rbpj. 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 Rbpj 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 Rbpj-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.