Mitochondrial Topo I CRISPR/Cas9 KO Plasmid (h): sc-408700
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Mitochondrial Topo I 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 Mitochondrial Topo I 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 | |
Mitochondrial Topo I CRISPR/Cas9 KO Plasmid (h) | sc-408700 | 20 µg | $397.00 |
Overview
TOP1MT encodes mitochondrial DNA topoisomerase I, a type IB topoisomerase that resolves torsional stress generated during mtDNA replication and transcription by introducing transient single-strand breaks and resealing them. By maintaining mitochondrial genome topology, TOP1MT supports mitochondrial gene expression, nucleoid organization, and oxidative phosphorylation capacity. Disruption of TOP1MT function has been linked to altered mtDNA integrity, impaired respiratory chain performance, and heightened sensitivity to mitochondrial stress. These processes are relevant to studies of mitochondrial dysfunction in neurodegeneration, cardiometabolic disease, and cancer cell bioenergetics.
Mitochondrial Topo I CRISPR/Cas9 KO Plasmid (h) is a pool of plasmids designed for targeted disruption of the TOP1MT gene in human cell lines. Each plasmid co-expresses a unique single guide RNA (sgRNA) targeting a distinct site within the TOP1MT 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 TOP1MT 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 Mitochondrial Topo I protein expression.
This CRISPR knockout system enables efficient generation of TOP1MT-deficient cell models for investigation of Mitochondrial Topo I signaling, functional genomics studies, cancer biology research, and evaluation of therapeutic responses in human cell lines.
Key Features
- sgRNAs targeting TOP1MT exon(s) critical for Mitochondrial Topo I 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 TOP1MT genomic sites to improve knockout efficiency
- Compatible with delivery by transfection
Design Variants
CRISPRs +/- HDRs
- gRNAs encoded by Mitochondrial Topo I CRISPR/Cas9 KO Plasmid (h) and Mitochondrial Topo I CRISPR/Cas9 KO Plasmid (h2) target distinct sites within the TOP1MT locus. One or both targeting designs may be available. See Related Products for availability.
- HDR donor constructs encoded by Mitochondrial Topo I HDR Plasmid (h) and Mitochondrial Topo I HDR Plasmid (h2) contain a puromycin resistance cassette and an RFP reporter flanked by TOP1MT homology arms to support homology-directed repair at defined TOP1MT 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.