neuroglobin CRISPR Activation Plasmid (h): sc-401696-ACT

Human NGB encodes neuroglobin, an oxygen-binding globin enriched in neurons that supports cellular adaptation to hypoxia and oxidative stress. Neuroglobin is implicated in maintaining mitochondrial function and redox homeostasis, with reported roles in modulating reactive oxygen species handling and nitric oxide–related signaling. Through these processes, NGB links oxygen availability to neuronal metabolism and stress-response pathways that influence survival under ischemic or inflammatory conditions. Altered neuroglobin expression has been associated with susceptibility to neurodegeneration and cerebrovascular injury models, making it a useful target for mechanistic studies of neuronal resilience.

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

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

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