VDAC3 CRISPR Activation Plasmid (h): sc-404240-ACT

VDAC3 (voltage-dependent anion-selective channel protein 3) is a mitochondrial outer membrane porin that contributes to metabolite and ion exchange between the cytosol and mitochondria, influencing oxidative phosphorylation and cellular bioenergetics. As part of the voltage-dependent anion channel family, it participates in regulating mitochondrial membrane permeability, redox balance, and cross-talk with apoptotic signaling pathways. VDAC3 has been linked to mitochondrial quality control processes and responses to oxidative stress, with particular relevance to tissues with high energetic demand. Dysregulated mitochondrial transport and signaling involving VDAC3 is of interest in studies of neurodegeneration, cardiometabolic dysfunction, infertility, and cancer-associated metabolic remodeling.

VDAC3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VDAC3 expression without altering the underlying DNA sequence.

VDAC3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VDAC3 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 VDAC3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous VDAC3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VDAC3 locus and enabling the study of VDAC3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of VDAC3 pathway restoration in tumor cells with silenced or reduced VDAC3 expression.

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