Nurr1 CRISPR Activation Plasmid (h): sc-400289-ACT

NR4A2 encodes the orphan nuclear receptor Nurr1, a transcription factor that regulates gene programs controlling neuronal differentiation, mitochondrial homeostasis, and anti-inflammatory signaling. Nurr1 is a key determinant of dopaminergic neuron development and maintenance, coordinating transcriptional networks involved in dopamine synthesis, vesicular transport, and stress response pathways. It interfaces with nuclear receptor signaling and chromatin-regulatory processes to modulate cell fate and survival under inflammatory or oxidative stress conditions. Dysregulated NR4A2/Nurr1 activity has been associated with neurodegeneration and altered immune activation, making it relevant for mechanistic studies of Parkinson’s disease-related pathways and neuroinflammatory phenotypes.

Nurr1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR4A2 expression without altering the underlying DNA sequence.

Nurr1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR4A2 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 NR4A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nurr1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR4A2 locus and enabling the study of Nurr1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nurr1 pathway restoration in tumor cells with silenced or reduced NR4A2 expression.

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