RNF182 Double Nickase Plasmid (h): sc-415138-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNF182 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
- RNF182 Double Nickase Plasmid (h) and RNF182 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RNF182. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RNF182 Double Nickase Plasmid (h) | sc-415138-NIC | 20 µg | $410.00 |
RNF182 encodes a RING finger E3 ubiquitin ligase that catalyzes ubiquitin transfer to specific protein substrates, thereby controlling protein stability and signaling amplitude in the ubiquitin–proteasome system. Through ubiquitination-dependent regulation of intracellular effectors, RNF182 is linked to pathways governing cellular stress responses, protein quality control, and turnover of membrane-associated and signaling proteins. Altered RNF182 expression has been reported in studies of nervous system biology and neuroinflammation, consistent with roles in neuronal homeostasis and regulated proteostasis. These features make RNF182 a useful target for investigating how ubiquitin ligases shape signaling networks and cell-state transitions in human cell models.
RNF182 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RNF182 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RNF182. 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 RNF182 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 RNF182-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.