α4a Tubulin Double Nickase Plasmid (h): sc-400016-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α4a 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
- α4a Tubulin Double Nickase Plasmid (h) and α4a Tubulin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TUBA4A. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: α4a Tubulin Antibody (MH-17): sc-134241
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
α4a Tubulin Double Nickase Plasmid (h) | sc-400016-NIC | 20 µg | $410.00 | |||
α4a Tubulin Double Nickase Plasmid (h2) | sc-400016-NIC-2 | 20 µg | $410.00 |
TUBA4A encodes α4a tubulin, a core α-tubulin isoform that heterodimerizes with β-tubulin to build microtubules required for cytoskeletal organization, intracellular transport, and mitotic spindle dynamics. Microtubule remodeling driven by tubulin isoform composition and post-translational modifications influences cell polarity, vesicle trafficking, and neurite outgrowth through pathways that coordinate motor proteins and microtubule-associated proteins. Altered tubulin homeostasis and microtubule dynamics are implicated in neurodegenerative phenotypes and axonal transport defects, and TUBA4A variation has been associated with motor neuron disease susceptibility in genetic studies. As a result, TUBA4A is commonly investigated in models of cytoskeletal stress responses, neuronal morphogenesis, and microtubule-targeting compound sensitivity.
α4a Tubulin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TUBA4A locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TUBA4A. 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 TUBA4A 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 TUBA4A-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.