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.
A20/TNFAIP3 Double Nickase Plasmid (h): sc-400447-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- A20/TNFAIP3 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
- A20/TNFAIP3 Double Nickase Plasmid (h) and A20/TNFAIP3 Double Nickase Plasmid (h2) 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 (h) | sc-400447-NIC | 20 µg | $410.00 | |||
A20/TNFAIP3 Double Nickase Plasmid (h2) | sc-400447-NIC-2 | 20 µg | $410.00 |
Human TNFAIP3 encodes A20, a ubiquitin-editing enzyme that acts as a key negative regulator of inflammatory signaling. A20 terminates NF-κB and MAPK pathway activation downstream of receptors such as TNFR, IL-1R, and TLRs by modulating K63- and M1-linked ubiquitin chains on signaling adaptors, thereby restricting cytokine production and preventing excessive cell stress responses. Through control of apoptosis and necroptosis checkpoints, A20 helps maintain immune homeostasis and tissue integrity. Dysregulation or loss-of-function variation in TNFAIP3 is linked to heightened inflammatory phenotypes and has been studied in contexts including autoimmunity, chronic inflammation, and tumor microenvironment signaling.
A20/TNFAIP3 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TNFAIP3 locus in human 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.