SUCLA2 CRISPR Activation Plasmid (h): sc-404356-ACT

Human SUCLA2 encodes the ATP-forming succinyl-CoA ligase subunit beta, a core mitochondrial matrix enzyme of the tricarboxylic acid (TCA) cycle that catalyzes the reversible conversion of succinyl-CoA to succinate with nucleotide phosphorylation. By linking carbon flux in oxidative metabolism to ATP/GTP generation, SUCLA2 supports mitochondrial bioenergetics, anaplerosis, and metabolic coupling to pathways such as heme biosynthesis and amino acid catabolism. SUCLA2 dysfunction is associated with mitochondrial DNA maintenance defects and impaired oxidative phosphorylation, contributing to neuromuscular and encephalomyopathic phenotypes reported in mitochondrial disease contexts. As a metabolic node, SUCLA2 is frequently studied in models of mitochondrial stress, redox imbalance, and energy-dependent cellular processes.

SUCLA2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SUCLA2 expression without altering the underlying DNA sequence.

SUCLA2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SUCLA2 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 SUCLA2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SUCLA2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SUCLA2 locus and enabling the study of SUCLA2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SUCLA2 pathway restoration in tumor cells with silenced or reduced SUCLA2 expression.

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