β1 Tubulin Double Nickase Plasmid (h): sc-400113-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- β1 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
- β1 Tubulin Double Nickase Plasmid (h) and β1 Tubulin Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TUBB1. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: β1 Tubulin Antibody (908G2X): sc-517652
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
β1 Tubulin Double Nickase Plasmid (h) | sc-400113-NIC | 20 µg | $410.00 | |||
β1 Tubulin Double Nickase Plasmid (h2) | sc-400113-NIC-2 | 20 µg | $410.00 |
TUBB1 encodes human β1 tubulin, a hematopoietic-lineage–enriched β-tubulin isotype that polymerizes with α-tubulins to form microtubules essential for cytoskeletal organization and intracellular transport. In megakaryocytes and platelets, β1 tubulin supports proplatelet formation, platelet shape change, and microtubule ring integrity, linking it to cytoskeletal dynamics, vesicle trafficking, and cell division processes. Perturbation of microtubule assembly or β1 tubulin expression can disrupt megakaryopoiesis and platelet biogenesis, and TUBB1 variants are associated with inherited platelet disorders and altered platelet function phenotypes. As a structural component of the microtubule network, β1 tubulin is also relevant to studies of mitotic spindle organization, cytoskeletal remodeling, and genotype–phenotype relationships in hematologic disease models.
β1 Tubulin Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TUBB1 locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TUBB1. 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 TUBB1 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 TUBB1-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.