RNF145 Double Nickase Plasmid (m): sc-428832-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNF145 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
- RNF145 Double Nickase Plasmid (m) and RNF145 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Rnf145. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RNF145 Double Nickase Plasmid (m) | sc-428832-NIC | 20 µg | $410.00 | |||
RNF145 Double Nickase Plasmid (m2) | sc-428832-NIC-2 | 20 µg | $410.00 |
Mouse Rnf145 encodes RNF145, an ER-resident RING-type E3 ubiquitin ligase that helps regulate cellular sterol and lipid homeostasis by promoting ubiquitin-dependent turnover of key pathway components. RNF145 activity interfaces with ER-associated protein quality control and proteostasis mechanisms, linking ubiquitination to adaptive responses in membrane lipid composition and metabolic state. Through its influence on cholesterol regulatory networks and related transcriptional programs, Rnf145 is frequently studied in the context of metabolic dysregulation and lipid-driven stress phenotypes. Loss- or gain-of-function interrogation of RNF145 supports mechanistic studies of ubiquitin signaling, ER membrane biology, and downstream effects on lipid metabolism pathways.
RNF145 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Rnf145 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Rnf145. 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 Rnf145 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 Rnf145-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.