ADH5 CRISPR Activation Plasmid (h): sc-403859-ACT

Human ADH5 encodes alcohol dehydrogenase 5 (also known as S-nitrosoglutathione reductase, GSNOR), an NAD-dependent dehydrogenase that governs cellular formaldehyde detoxification and nitric oxide bioactivity by metabolizing S-nitrosoglutathione. Through regulation of protein S-nitrosylation and redox balance, ADH5 influences stress response signaling, mitochondrial function, and pathways linked to inflammation and DNA damage. Altered ADH5 activity has been associated with dysregulated nitrosative stress and impaired aldehyde clearance, processes implicated in cardiopulmonary biology, immune signaling, and cancer-relevant redox adaptation. As a result, ADH5 is frequently studied in models of oxidative/nitrosative injury, metabolism-driven signaling, and genome stability under aldehyde stress.

ADH5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ADH5 expression without altering the underlying DNA sequence.

ADH5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ADH5 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 ADH5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ADH5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ADH5 locus and enabling the study of ADH5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ADH5 pathway restoration in tumor cells with silenced or reduced ADH5 expression.

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