Topo IIα CRISPR Activation Plasmid (h): sc-400869-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Topo IIα CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Topo IIα 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 IIα CRISPR Activation Plasmid (h) and Topo IIα CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the TOP2A transcriptional start site. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: Topo IIα Antibody (G-6): sc-166934
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Topo IIα CRISPR Activation Plasmid (h) | sc-400869-ACT | 20 µg | $397.00 | |||
Topo IIα CRISPR Activation Plasmid (h2) | sc-400869-ACT-2 | 20 µg | $397.00 |
TOP2A encodes human DNA topoisomerase IIα, an ATP-dependent enzyme that resolves DNA supercoiling and DNA catenanes by generating transient double-strand breaks, enabling faithful chromosome condensation and segregation. Topo IIα is a central component of DNA replication and transcriptional stress responses and is required for decatenation during late S phase and G2/M progression. Its activity is tightly linked to cell cycle regulation, DNA damage signaling, and maintenance of genome stability. Dysregulated TOP2A expression is frequently associated with highly proliferative states and is studied in the context of chromosomal instability and cancer-related biology.
Topo IIα CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous TOP2A expression without altering the underlying DNA sequence.
Topo IIα CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the TOP2A 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 TOP2A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Topo IIα expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native TOP2A locus and enabling the study of Topo IIα-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Topo IIα pathway restoration in tumor cells with silenced or reduced TOP2A expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.