SDHD CRISPR Activation Plasmid (h): sc-403276-ACT

Human SDHD encodes the small integral membrane subunit D of succinate dehydrogenase (mitochondrial complex II), a core component that anchors the SDH catalytic domain within the inner mitochondrial membrane and supports electron transfer from succinate to ubiquinone. By linking the tricarboxylic acid (TCA) cycle to the respiratory chain, SDHD contributes to oxidative phosphorylation, cellular redox balance, and metabolite homeostasis, including regulation of succinate levels. Disruption or altered expression of SDHD can promote mitochondrial dysfunction and succinate accumulation, reshaping signaling through oxygen-sensing and stress-response pathways such as HIF-related transcriptional programs. SDHD has strong disease relevance in SDH-deficient tumor biology and mitochondrial metabolic dysregulation, making it a key target for studying genotype–metabolism relationships.

SDHD CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SDHD expression without altering the underlying DNA sequence.

SDHD CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SDHD 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 SDHD transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SDHD expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SDHD locus and enabling the study of SDHD-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SDHD pathway restoration in tumor cells with silenced or reduced SDHD expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.