α-KGD CRISPR Activation Plasmid (h): sc-402915-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- α-KGD CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- α-KGD 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 α-KGD CRISPR Activation Plasmid (h) and α-KGD CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the OGDH transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
α-KGD CRISPR Activation Plasmid (h) | sc-402915-ACT | 20 µg | $397.00 | |||
α-KGD CRISPR Activation Plasmid (h2) | sc-402915-ACT-2 | 20 µg | $397.00 |
OGDH encodes the E1 component of the mitochondrial 2-oxoglutarate dehydrogenase complex (α-KGD), a rate-limiting enzyme in the tricarboxylic acid (TCA) cycle that converts α-ketoglutarate to succinyl-CoA while generating NADH for oxidative phosphorylation. Through control of α-ketoglutarate flux, OGDH links central carbon metabolism to amino acid catabolism, redox balance, and mitochondrial reactive oxygen species handling. Changes in OGDH activity can alter levels of α-ketoglutarate and related metabolites that influence epigenetic regulation via dioxygenases, connecting mitochondrial metabolism to chromatin state. Dysregulation of this node has been associated with mitochondrial metabolic dysfunction and has been studied in contexts including neurodegeneration and cancer metabolism.
α-KGD CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous OGDH expression without altering the underlying DNA sequence.
α-KGD CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the OGDH 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 OGDH transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous α-KGD expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native OGDH locus and enabling the study of α-KGD-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of α-KGD pathway restoration in tumor cells with silenced or reduced OGDH expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.