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 IIIβ CRISPR/Cas9 KO Plasmid (m): sc-423472
- Target species: mouse
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIIβ CRISPR/Cas9 Knockout (KO) Plasmid (m) 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 IIIβ 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
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Topo IIIβ-1 Antibody (B-10): sc-137238
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Topo IIIβ CRISPR/Cas9 KO Plasmid (m) | sc-423472 | 20 µg | $397.00 |
Overview
Top3b encodes DNA topoisomerase III beta (Topo IIIβ), a type IA topoisomerase that resolves topological stress and recombination intermediates during genome maintenance. In mouse cells, Topo IIIβ is implicated in preserving nucleic acid integrity through roles in DNA repair, replication-associated stress responses, and regulation of RNA metabolism, consistent with its ability to act on both DNA and RNA substrates. These activities intersect with pathways controlling transcription-replication conflicts and recombination-driven genome stability, processes that influence cellular homeostasis. Disruption of Top3b-dependent nucleic acid processing has been associated with genomic instability phenotypes and has relevance to studies of neurodevelopmental and cancer-related mechanisms in model systems.
Topo IIIβ CRISPR/Cas9 KO Plasmid (m) is a pool of plasmids designed for targeted disruption of the Top3b gene in mouse cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the Top3b 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 Top3b 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 IIIβ protein expression.
This CRISPR knockout system enables efficient generation of Top3b-deficient cell models for investigation of Topo IIIβ signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting Top3b exon(s) critical for Topo IIIβ 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 Top3b genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Topo IIIβ CRISPR/Cas9 KO Plasmid (m) and Topo IIIβ CRISPR/Cas9 KO Plasmid (m2) target distinct sites within the Top3b locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Topo IIIβ HDR Plasmid (m) and Topo IIIβ HDR Plasmid (m2) contain a puromycin resistance cassette and an RFP reporter flanked by Top3b homology arms to support homology-directed repair at defined Top3b 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.