DGLUCY CRISPR Activation Plasmid (h): sc-413322-ACT

C14orf159 encodes DGLUCY, a mitochondrial protein implicated in mitochondrial gene expression and respiratory chain homeostasis. Emerging functional studies associate DGLUCY with regulation of oxidative phosphorylation efficiency, mitochondrial ribosome-linked processes, and cellular responses to metabolic stress. Altered DGLUCY activity has been discussed in the context of mitochondrial dysfunction phenotypes, where shifts in bioenergetic capacity can influence redox balance and susceptibility to stress. As a result, DGLUCY is of interest for mechanistic research on mitochondrial metabolism, quality control pathways, and disease-relevant states characterized by impaired mitochondrial function.

DGLUCY CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous C14orf159 expression without altering the underlying DNA sequence.

DGLUCY CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the C14orf159 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 C14orf159 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous DGLUCY expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native C14orf159 locus and enabling the study of DGLUCY-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of DGLUCY pathway restoration in tumor cells with silenced or reduced C14orf159 expression.

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