Nrf1 CRISPR Activation Plasmid (m): sc-421868-ACT

Mouse Nfe2l1 encodes the CNC-bZIP transcription factor Nrf1, a key regulator of cellular proteostasis and redox balance. Nrf1 coordinates transcriptional programs controlling proteasome subunit biogenesis and ubiquitin-dependent protein turnover, linking endoplasmic reticulum homeostasis to antioxidant and metabolic stress responses. Through crosstalk with NRF2 signaling and control of detoxification and mitochondrial-associated pathways, Nrf1 helps maintain cellular resilience under proteotoxic stress. Dysregulation of Nfe2l1/Nrf1 activity has been associated with altered proteasome capacity, oxidative damage, and metabolic dysfunction, making it relevant to mechanistic studies of neurodegeneration, liver biology, and cancer cell stress adaptation.

Nrf1 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nfe2l1 expression without altering the underlying DNA sequence.

Nrf1 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nfe2l1 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 Nfe2l1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nrf1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nfe2l1 locus and enabling the study of Nrf1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nrf1 pathway restoration in tumor cells with silenced or reduced Nfe2l1 expression.

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