RNF25 Double Nickase Plasmid (h): sc-408040-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- RNF25 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
- RNF25 Double Nickase Plasmid (h) and RNF25 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting RNF25. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: RNF25 Antibody (E-10): sc-398749
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
RNF25 Double Nickase Plasmid (h) | sc-408040-NIC | 20 µg | $410.00 | |||
RNF25 Double Nickase Plasmid (h2) | sc-408040-NIC-2 | 20 µg | $410.00 |
RNF25 encodes an E3 ubiquitin-protein ligase containing a RING finger domain that helps regulate protein stability and signaling by catalyzing ubiquitin transfer to specific substrates. Through ubiquitin-dependent control of protein turnover and signaling complex composition, RNF25 can influence pathways linked to innate immune and inflammatory responses, including modulation of NF-κB–associated transcriptional programs. As a component of the cellular ubiquitin–proteasome system, RNF25 contributes to proteostasis and signal termination, processes frequently perturbed in cancer biology and immune dysregulation. Altered expression or activity of ubiquitin ligases such as RNF25 is therefore relevant to studies of stress signaling, cytokine-driven transcription, and pathway cross-talk in human cells.
RNF25 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the RNF25 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within RNF25. 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 RNF25 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 RNF25-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.