RNF145 Double Nickase Plasmid (h): sc-416912-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNF145 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
- RNF145 Double Nickase Plasmid (h) and RNF145 Double Nickase Plasmid (h2) 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 (h) | sc-416912-NIC | 20 µg | $410.00 | |||
RNF145 Double Nickase Plasmid (h2) | sc-416912-NIC-2 | 20 µg | $410.00 |
RNF145 encodes a multi-pass endoplasmic reticulum membrane RING finger E3 ubiquitin ligase that helps control cellular cholesterol and lipid homeostasis by regulating the stability of key biosynthetic enzymes. As an ER-associated ubiquitination factor, RNF145 integrates with the ubiquitin–proteasome system and sterol-responsive feedback programs to fine-tune mevalonate pathway output and membrane lipid composition. This regulation supports broader processes including ER quality control, metabolic adaptation, and stress signaling linked to proteostasis. Dysregulated sterol metabolism and ER proteostasis are commonly implicated in cardiometabolic and proliferative disease biology, making RNF145 a useful target for mechanistic studies of lipid-driven phenotypes.
RNF145 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RNF145 locus in human 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.