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.
GAPDH CRISPR Activation Plasmid (h): sc-400002-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- GAPDH CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- GAPDH 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 GAPDH CRISPR Activation Plasmid (h) and GAPDH CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the GAPDH 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: GAPDH Antibody (0411): sc-47724
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
GAPDH CRISPR Activation Plasmid (h) | sc-400002-ACT | 20 µg | $397.00 | |||
GAPDH CRISPR Activation Plasmid (h2) | sc-400002-ACT-2 | 20 µg | $397.00 |
Human GAPDH encodes glyceraldehyde-3-phosphate dehydrogenase, a central glycolytic enzyme that catalyzes the conversion of glyceraldehyde-3-phosphate to 1,3-bisphosphoglycerate while generating NADH. Beyond energy metabolism, GAPDH participates in redox homeostasis, RNA binding, membrane trafficking, and stress-responsive signaling, with context-dependent roles in apoptosis and autophagy. Its expression and post-translational regulation track with metabolic reprogramming and hypoxia-responsive pathways, making it relevant to studies of proliferative states, inflammation, and neurodegenerative processes where glycolytic flux and oxidative stress are altered.
GAPDH CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous GAPDH expression without altering the underlying DNA sequence.
GAPDH CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the GAPDH 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 GAPDH transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous GAPDH expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native GAPDH locus and enabling the study of GAPDH-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of GAPDH pathway restoration in tumor cells with silenced or reduced GAPDH expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.