Ring1b Double Nickase Plasmid (h): sc-404633-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Ring1b 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
- Ring1b Double Nickase Plasmid (h) and Ring1b Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RNF2. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RING1B Antibody (N-32): sc-101109
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Ring1b Double Nickase Plasmid (h) | sc-404633-NIC | 20 µg | $410.00 | |||
Ring1b Double Nickase Plasmid (h2) | sc-404633-NIC-2 | 20 µg | $410.00 |
RNF2 encodes Ring1b, a RING finger E3 ubiquitin ligase that forms the catalytic core of Polycomb repressive complex 1 (PRC1). Ring1b monoubiquitinates histone H2A at Lys119, reinforcing Polycomb-mediated chromatin compaction and transcriptional repression during development and lineage specification. Through coordination with PRC2-dependent H3K27 methylation and other epigenetic regulators, RNF2 helps control cell-cycle programs, differentiation states, and DNA damage responses. Dysregulated RNF2 activity and PRC1 signaling are implicated in altered transcriptional networks observed across multiple cancer types and other disorders linked to epigenetic misregulation.
Ring1b Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RNF2 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RNF2. 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 RNF2 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 RNF2-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.