NDUFB5 CRISPR Activation Plasmid (h): sc-410902-ACT

Human NDUFB5 encodes a small accessory subunit of mitochondrial respiratory chain complex I (NADH:ubiquinone oxidoreductase) that supports proper assembly and electron transfer within oxidative phosphorylation. By contributing to proton pumping and maintenance of mitochondrial membrane potential, NDUFB5 influences ATP production and redox homeostasis, processes tightly linked to ROS signaling and metabolic adaptation. Altered complex I function is broadly relevant to mitochondrial disease biology and has been observed across contexts of neuromuscular dysfunction, neurodegeneration, and tumor metabolism where bioenergetic stress and mitochondrial remodeling are prominent. As such, NDUFB5 is a useful target for studying respiratory chain integrity, mito-nuclear communication, and pathways that couple energy metabolism to cell fate decisions.

NDUFB5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NDUFB5 expression without altering the underlying DNA sequence.

NDUFB5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NDUFB5 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 NDUFB5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NDUFB5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NDUFB5 locus and enabling the study of NDUFB5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NDUFB5 pathway restoration in tumor cells with silenced or reduced NDUFB5 expression.

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