Topo IIIα CRISPR Activation Plasmid (h): sc-404439-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIIα CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Topo IIIα CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by Topo IIIα CRISPR Activation Plasmid (h) and Topo IIIα CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the TOP3A transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Topo IIIα CRISPR Activation Plasmid (h) | sc-404439-ACT | 20 µg | $397.00 |
TOP3A encodes DNA topoisomerase IIIα, a type IA topoisomerase that resolves DNA topological stress and recombination intermediates during replication and repair. Topo IIIα functions with the BLM helicase, RMI1, and RMI2 in the BTRR complex to dissolve double Holliday junctions and suppress aberrant crossover events, thereby promoting genome stability. Through roles in homologous recombination, replication fork restart, and decatenation of entangled DNA, TOP3A helps maintain proper chromosome segregation and mitochondrial DNA maintenance. Disruption of this network is linked to elevated sister chromatid exchanges and genomic instability phenotypes relevant to cancer biology and inherited genome maintenance disorders.
Topo IIIα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TOP3A expression without altering the underlying DNA sequence.
Topo IIIα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TOP3A locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.
Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the TOP3A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Topo IIIα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TOP3A locus and enabling the study of Topo IIIα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Topo IIIα pathway restoration in tumor cells with silenced or reduced TOP3A expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.