RNF149 Double Nickase Plasmid (m): sc-426694-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNF149 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
- RNF149 Double Nickase Plasmid (m) and RNF149 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Rnf149. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RNF149 Double Nickase Plasmid (m) | sc-426694-NIC | 20 µg | $410.00 |
Mouse Rnf149 encodes RNF149, a RING finger E3 ubiquitin ligase that contributes to ubiquitin-dependent protein turnover and regulation of cellular proteostasis. As part of ubiquitin–proteasome system control, RNF149 is relevant to pathways governing protein quality control, signaling amplitude, and stress-adaptive responses that shape cell survival and homeostasis. Altered ubiquitination programs are broadly implicated in disease-associated phenotypes, including dysregulated growth control and neurodegeneration-linked proteotoxic stress, making RNF149 a useful node for mechanistic studies. Functional interrogation of RNF149 supports research into how E3 ligases tune substrate stability and downstream pathway outputs in mammalian cells.
RNF149 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Rnf149 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Rnf149. 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 Rnf149 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 Rnf149-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.