NDUFV1 CRISPR Activation Plasmid (h): sc-404372-ACT

NDUFV1 encodes a core subunit of mitochondrial complex I (NADH:ubiquinone oxidoreductase) that participates in electron transfer from NADH to ubiquinone, supporting oxidative phosphorylation and cellular ATP production. As part of the respiratory chain, NDUFV1 contributes to maintenance of mitochondrial membrane potential and redox homeostasis, linking mitochondrial metabolism to reactive oxygen species signaling. Altered complex I function is associated with mitochondrial dysfunction phenotypes and has been implicated in neurometabolic and neurodegenerative disease mechanisms. NDUFV1 is therefore widely studied in pathways governing energy metabolism, mitochondrial quality control, and stress responses.

NDUFV1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NDUFV1 expression without altering the underlying DNA sequence.

NDUFV1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NDUFV1 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 NDUFV1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NDUFV1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NDUFV1 locus and enabling the study of NDUFV1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NDUFV1 pathway restoration in tumor cells with silenced or reduced NDUFV1 expression.

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