Fatso CRISPR Activation Plasmid (h): sc-403708-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- Fatso CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- Fatso 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 Fatso CRISPR Activation Plasmid (h) and Fatso CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the FTO 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: Fatso Antibody (C-3): sc-271713
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
Fatso CRISPR Activation Plasmid (h) | sc-403708-ACT | 20 µg | $397.00 |
Human FTO (Fatso) encodes an Fe(II)/2-oxoglutarate–dependent RNA demethylase that catalyzes removal of N6-methyladenosine (m6A) and related methyl marks, thereby tuning mRNA stability, splicing, and translation. Through m6A-dependent control of transcript fate, FTO influences nutrient sensing, adipogenesis, and energy homeostasis programs, integrating with pathways that regulate cellular metabolism and stress responses. Altered FTO expression or activity has been linked to metabolic phenotypes and obesity-associated traits, and it is also studied in contexts where RNA epitranscriptomic regulation shapes cell-state transitions. As a result, FTO is widely used as a node for probing how RNA methylation impacts gene regulatory networks and metabolic remodeling in human cells.
Fatso CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous FTO expression without altering the underlying DNA sequence.
Fatso CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the FTO 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 FTO transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Fatso expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native FTO locus and enabling the study of Fatso-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Fatso pathway restoration in tumor cells with silenced or reduced FTO expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.