β2A Tubulin Double Nickase Plasmid (h): sc-400054-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- β2A Tubulin 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
- β2A Tubulin Double Nickase Plasmid (h) and β2A Tubulin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TUBB2A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: β2A Tubulin Antibody (2-RY22): sc-134229
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
β2A Tubulin Double Nickase Plasmid (h) | sc-400054-NIC | 20 µg | $410.00 | |||
β2A Tubulin Double Nickase Plasmid (h2) | sc-400054-NIC-2 | 20 µg | $410.00 |
TUBB2A encodes human β2A tubulin, a core component of α/β-tubulin heterodimers that polymerize into microtubules to support cytoskeletal architecture, intracellular transport, and mitotic spindle assembly. Microtubule dynamics coordinate processes spanning vesicle trafficking, cell polarity, and chromosome segregation, integrating with cell-cycle control and neuronal morphogenesis pathways. Altered β-tubulin composition or microtubule instability can disrupt neurodevelopmental programs and has been associated with tubulinopathy phenotypes, including cortical malformations and epilepsy-related presentations. As a highly conserved cytoskeletal protein, β2A tubulin is also used to interrogate mechanisms of microtubule-dependent signaling and stress responses in human cells.
β2A Tubulin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TUBB2A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TUBB2A. 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 TUBB2A 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 TUBB2A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.