NRF-1 CRISPR Activation Plasmid (h2): sc-400938-ACT-2

Human NRF1 encodes nuclear respiratory factor 1 (NRF-1), a sequence-specific transcription factor that coordinates expression of nuclear-encoded mitochondrial genes, including components of the electron transport chain, mitochondrial transcription/replication machinery (e.g., TFAM), and factors supporting oxidative phosphorylation and mitochondrial biogenesis. NRF-1 integrates cellular energy and redox cues with cell growth programs by regulating transcriptional networks linked to proteostasis, heme biosynthesis, and mitochondrial–nuclear communication, thereby influencing metabolic remodeling and stress adaptation. Dysregulated NRF1/NRF-1 activity has been associated with mitochondrial dysfunction and altered bioenergetics observed across neurodegeneration, cardiometabolic disorders, and cancer contexts. Gene editing or perturbation of NRF1 enables mechanistic studies of mitochondrial gene regulation, respiratory capacity, and transcriptional circuitry in human cell models, including pathway mapping and genotype–phenotype analyses using omics readouts.

NRF-1 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous NRF1 expression without altering the underlying DNA sequence.

NRF-1 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NRF1 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 NRF1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NRF-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NRF1 locus and enabling the study of NRF-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NRF-1 pathway restoration in tumor cells with silenced or reduced NRF1 expression.

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