A20/TNFAIP3 Double Nickase Plasmid (m): sc-423436-NIC
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- A20/TNFAIP3 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
- A20/TNFAIP3 Double Nickase Plasmid (m) and A20/TNFAIP3 Double Nickase Plasmid (m2) encode distinct paired gRNA designs targeting Tnfaip3. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: A20/TNFAIP3 Antibody (A-12): sc-166692
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
A20/TNFAIP3 Double Nickase Plasmid (m) | sc-423436-NIC | 20 µg | $410.00 | |||
A20/TNFAIP3 Double Nickase Plasmid (m2) | sc-423436-NIC-2 | 20 µg | $410.00 |
Mouse Tnfaip3 encodes A20/TNFAIP3, a ubiquitin-editing enzyme that functions as a central negative regulator of inflammatory signaling. A20 restricts NF-κB and MAPK pathway activation downstream of receptors such as TNFR, IL-1R, and Toll-like receptors by modulating K63- and M1-linked ubiquitin chains on key signaling adaptors. Through control of cytokine-driven transcriptional programs, A20 influences innate and adaptive immune homeostasis, cell survival, and stress responses. Dysregulated A20 activity is implicated in models of autoimmunity, chronic inflammation, and oncogenic signaling where persistent NF-κB activation contributes to disease-relevant phenotypes.
A20/TNFAIP3 Double Nickase Plasmid (m) consists of a matched pair of plasmids engineered for high-specificity editing of the Tnfaip3 locus in mouse cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within Tnfaip3. 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 Tnfaip3 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 Tnfaip3-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.