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.
ADO CRISPR Activation Plasmid (h2): sc-408985-ACT-2
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- ADO CRISPR Activation Plasmid (h2) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- ADO CRISPR Activation Plasmid (h2) 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 ADO CRISPR Activation Plasmid (h2) and ADO CRISPR Activation Plasmid (h22) target distinct regulatory regions upstream of the ADO 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: ADO Antibody (E-11): sc-515318
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
ADO CRISPR Activation Plasmid (h2) | sc-408985-ACT-2 | 20 µg | $397.00 |
Human ADO (2-aminoethanethiol dioxygenase) is a non-heme iron enzyme that catalyzes the oxygen-dependent oxidation of cysteamine to hypotaurine, supporting cellular taurine biosynthesis and broader sulfur amino acid metabolism. By linking cysteamine turnover to redox and mitochondrial metabolic homeostasis, ADO activity intersects with pathways regulating oxidative stress responses and bioenergetic adaptation in metabolically active tissues. Dysregulation of taurine/hypotaurine balance and thiol metabolism has been implicated in conditions associated with mitochondrial dysfunction and altered reactive oxygen species handling, making ADO a useful target for mechanistic studies. Gene editing or perturbation of ADO enables researchers to interrogate cysteamine-derived metabolite flux, iron-dependent dioxygenase biology, and downstream impacts on cellular stress phenotypes in relevant human model systems.
ADO CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous ADO expression without altering the underlying DNA sequence.
ADO CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADO 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 ADO transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADO expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADO locus and enabling the study of ADO-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADO pathway restoration in tumor cells with silenced or reduced ADO expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.