TNF alpha Double Nickase Plasmid (h): sc-400124-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TNF alpha 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
- TNF alpha Double Nickase Plasmid (h) and TNF alpha Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TNF. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TNF alpha Antibody (C-4): sc-133192
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TNF alpha Double Nickase Plasmid (h) | sc-400124-NIC | 20 µg | $410.00 | |||
TNF alpha Double Nickase Plasmid (h2) | sc-400124-NIC-2 | 20 µg | $410.00 |
TNF encodes tumor necrosis factor alpha (TNFα), a pleiotropic pro-inflammatory cytokine produced primarily by activated macrophages and T cells that coordinates innate and adaptive immune responses. TNFα signals through TNFR1 and TNFR2 to regulate canonical NF-κB and MAPK cascades, while also influencing apoptosis and necroptosis depending on receptor context and downstream adaptor engagement. These pathways shape cytokine networks, leukocyte recruitment, endothelial activation, and tissue remodeling in response to infection or injury. Dysregulated TNFα activity is widely used as a molecular readout in studies of chronic inflammation and immune-mediated pathology, including autoimmunity, inflammatory bowel disease biology, and tumor microenvironment signaling.
TNF alpha Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TNF locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TNF. 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 TNF 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 TNF-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.