Topo IIIα CRISPR/Cas9 KO Plasmid (h): sc-404439
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIIα CRISPR/Cas9 Knockout (KO) Plasmid (h) is a pool of plasmids, each encoding Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed for maximum knockout efficiency using sequences derived from the GeCKO v2 library
- gRNA sequences direct Cas9 to induce site-specific double-strand breaks (DSBs) in the Topo IIIα genomic locus, resulting in gene knockout through non-homologous end joining (NHEJ)
- The puromycin resistance and RFP genes are flanked by LoxP sites, enabling removal of selection markers via Cre recombinase (Cre Vector: sc-418923) after establishing stable knockout cell lines
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Topo IIIα CRISPR/Cas9 KO Plasmid (h) | sc-404439 | 20 µg | $397.00 |
Overview
TOP3A encodes human DNA topoisomerase IIIα, a type IA topoisomerase that resolves single-stranded DNA topological intermediates during DNA replication, recombination, and repair. Topo IIIα functions in the BTRR complex with BLM, RMI1, and RMI2 to dissolve double Holliday junctions, suppress aberrant crossover events, and maintain genome stability during homologous recombination. It also supports mitochondrial DNA maintenance through cooperation with organelle-specific replication and repair factors, linking TOP3A to nuclear and mitochondrial genome integrity. Disruption of TOP3A-associated pathways is relevant to genome instability phenotypes and has been connected to inherited disorders featuring defective DNA repair and replication stress sensitivity.
Topo IIIα CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TOP3A gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TOP3A together with the Streptococcus pyogenes Cas9 nuclease. The plasmids also encode GFP, allowing fluorescent identification and enrichment of successfully transfected cells by fluorescence microscopy or flow cytometry.
The multi-guide design increases the likelihood of generating insertions or deletions (indels) that disrupt the TOP3A open reading frame following Cas9-mediated double-strand break formation. DNA breaks introduced by the CRISPR/Cas9 system are repaired through endogenous non-homologous end joining (NHEJ) pathways, frequently resulting in frameshift mutations that abolish Topo IIIα protein expression.
This CRISPR knockout system enables efficient generation of TOP3A-deficient cell models for investigation of Topo IIIα signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TOP3A exon(s) critical for Topo IIIα function
- Co-expression of SpCas9 and sgRNA from a single plasmid for simplified delivery
- GFP reporter for identification of transfected cells
- Pool of plasmids targeting multiple TOP3A genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Topo IIIα CRISPR/Cas9 KO Plasmid (h) and Topo IIIα CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TOP3A locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Topo IIIα HDR Plasmid (h) and Topo IIIα HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TOP3A homology arms to support homology-directed repair at defined TOP3A target sites corresponding to the CRISPR/Cas9 KO designs. HDR donor availability may vary. See Related Products for availability.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.