TNFβ Double Nickase Plasmid (h): sc-403005-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- TNFβ 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
- TNFβ Double Nickase Plasmid (h) and TNFβ Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting LTA. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: TNFβ Antibody (E-6): sc-28345
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
TNFβ Double Nickase Plasmid (h) | sc-403005-NIC | 20 µg | $410.00 | |||
TNFβ Double Nickase Plasmid (h2) | sc-403005-NIC-2 | 20 µg | $410.00 |
Human LTA encodes lymphotoxin-α (TNFβ), a TNF superfamily cytokine produced primarily by activated lymphocytes that signals through TNFR1/TNFR2 and, in membrane-associated complexes with LTβ, via LTβR. TNFβ engages canonical NF-κB and MAPK pathways to regulate inflammatory gene expression, cell survival and apoptosis, and the organization of lymphoid tissue microenvironments. It contributes to immune cell–stromal crosstalk, endothelial activation, and cytokine networks that shape adaptive immune responses. Dysregulated LTA signaling has been linked to chronic inflammation and immune-mediated pathologies, as well as tumor–immune interactions in the microenvironment.
TNFβ Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the LTA locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within LTA. 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 LTA 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 LTA-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.