SDHA CRISPR Activation Plasmid (h): sc-401689-ACT

SDHA encodes the succinate dehydrogenase complex flavoprotein subunit A, a core catalytic component of mitochondrial complex II that links the tricarboxylic acid cycle to the electron transport chain by oxidizing succinate to fumarate and transferring electrons to ubiquinone. Through its role in oxidative phosphorylation and redox homeostasis, SDHA influences cellular ATP production, mitochondrial metabolism, and reactive oxygen species balance. Altered SDHA function can disrupt bioenergetic flux and metabolite signaling, with downstream effects on hypoxia response and mitochondrial stress pathways. Genetic and functional perturbations of SDHA have been associated with mitochondrial disease phenotypes and SDH-deficient tumor biology, supporting its utility as a model target in metabolic and cancer research.

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

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

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