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.
Topo IIβ Double Nickase Plasmid (h): sc-400773-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIβ 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
- Topo IIβ Double Nickase Plasmid (h) and Topo IIβ Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting TOP2B. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Topo IIβ Antibody (A-12): sc-365071
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Topo IIβ Double Nickase Plasmid (h) | sc-400773-NIC | 20 µg | $410.00 | |||
Topo IIβ Double Nickase Plasmid (h2) | sc-400773-NIC-2 | 20 µg | $410.00 |
TOP2B encodes human DNA topoisomerase IIβ (Topo IIβ), an ATP-dependent enzyme that resolves DNA supercoiling and catenanes by generating transient double-strand breaks and re-ligating DNA. Topo IIβ is important for transcriptional regulation, chromatin topology, and proper progression through DNA replication and chromosome segregation, linking it to DNA damage response and genome stability pathways. Its activity intersects with processes such as sister chromatid decatenation, torsional stress relief during transcription, and maintenance of higher-order chromatin structure. Dysregulation of TOP2B function or aberrant topoisomerase II–mediated DNA breaks has been associated with genomic instability and altered gene expression programs studied in cancer biology and neurodevelopmental models.
Topo IIβ Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the TOP2B locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within TOP2B. 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 TOP2B 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 TOP2B-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.