T-bet/TBX21 Double Nickase Plasmid (h): sc-400480-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- T-bet/TBX21 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
- T-bet/TBX21 Double Nickase Plasmid (h) and T-bet/TBX21 Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TBX21. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: T-bet/TBX21 Antibody (4B10): sc-21749
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
T-bet/TBX21 Double Nickase Plasmid (h) | sc-400480-NIC | 20 µg | $410.00 | |||
T-bet/TBX21 Double Nickase Plasmid (h2) | sc-400480-NIC-2 | 20 µg | $410.00 |
TBX21 encodes T-bet, a T-box transcription factor that orchestrates type 1 immune programs by promoting IFNG expression and reinforcing Th1 lineage commitment while antagonizing alternative T helper fates. T-bet integrates signals downstream of cytokine and antigen receptor pathways, including IL-12/STAT4 and IFN-γ/STAT1, to shape chromatin accessibility and transcriptional networks in CD4+ and CD8+ T cells as well as NK cells. Through control of effector differentiation, cytotoxic function, and inflammatory cytokine balance, TBX21 is widely studied in contexts of chronic infection, tumor immunobiology, and immune-mediated inflammatory disease. Dysregulated TBX21 activity or expression has been linked to aberrant immune polarization and altered tissue inflammation, making it a useful node for mechanistic studies of immune regulation.
T-bet/TBX21 Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TBX21 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TBX21. 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 TBX21 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 TBX21-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.