Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
EFP Double Nickase Plasmid (m): sc-432163-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- EFP 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
- EFP Double Nickase Plasmid (m) and EFP Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Trim25. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: EFP Antibody (E-4): sc-166926
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
EFP Double Nickase Plasmid (m) | sc-432163-NIC | 20 µg | $410.00 | |||
EFP Double Nickase Plasmid (m2) | sc-432163-NIC-2 | 20 µg | $410.00 |
Mouse Trim25 encodes EFP (TRIM25), a RING-type E3 ubiquitin ligase that regulates protein stability and signaling through ubiquitin-dependent post-translational modification. EFP participates in innate immune and inflammatory pathways by modulating pattern-recognition receptor signaling and interferon-stimulated responses, and it also influences RNA metabolism through interactions with RNA-binding proteins. In addition to antiviral defense, TRIM25 has been linked to regulation of cell cycle and stress responses, making it relevant for studying context-dependent control of transcriptional programs and proteostasis. Altered TRIM25 activity and ubiquitination networks are associated with immune dysregulation and cancer-related phenotypes, supporting its use as a mechanistic node in pathway-centric research.
EFP Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Trim25 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Trim25. 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 Trim25 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 Trim25-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.