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.
brachyury Double Nickase Plasmid (h): sc-416539-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- brachyury 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
- brachyury Double Nickase Plasmid (h) and brachyury Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting T. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: brachyury Antibody (D-10): sc-166962
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
brachyury Double Nickase Plasmid (h) | sc-416539-NIC | 20 µg | $410.00 | |||
brachyury Double Nickase Plasmid (h2) | sc-416539-NIC-2 | 20 µg | $410.00 |
Human T encodes the T-box transcription factor brachyury, a master regulator of mesoderm specification and posterior body axis formation during early development. Brachyury binds T-box DNA motifs to control gene programs that coordinate epithelial–mesenchymal transition, cell migration, and lineage commitment, integrating with WNT/β-catenin, FGF, and TGF-β signaling networks. In adult and disease contexts, altered brachyury activity is linked to aberrant differentiation states and invasiveness-associated transcriptional programs, making it a widely used marker and mechanistic node in developmental biology and tumor cell plasticity research. Dissecting T-dependent transcriptional regulation supports studies of fate decisions, chromatin state dynamics, and signaling cross-talk in model systems and engineered cell lines.
brachyury Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the T locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within T. 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 T 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 T-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.