SDHB CRISPR Activation Plasmid (h): sc-401529-ACT

SDHB encodes the iron–sulfur subunit of succinate dehydrogenase (mitochondrial complex II), a dual-function enzyme that couples the tricarboxylic acid (TCA) cycle to the electron transport chain by catalyzing succinate oxidation and transferring electrons to ubiquinone. By sustaining mitochondrial respiration and redox balance, SDHB influences oxidative phosphorylation, reactive oxygen species signaling, and cellular metabolic state. Perturbation of SDHB function is linked to metabolic reprogramming and altered hypoxia signaling, and germline or somatic changes in SDHB are associated with susceptibility to paraganglioma/pheochromocytoma and related mitochondrial dysfunction phenotypes.

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

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

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