Nurr1 CRISPR Activation Plasmid (r): sc-437375-ACT

Nurr1 (NR4A2) is an orphan nuclear receptor transcription factor that regulates gene programs required for dopaminergic neuron differentiation, survival, and synaptic function in the rat nervous system. It coordinates activity-dependent transcription and interacts with pathways controlling mitochondrial homeostasis, oxidative stress responses, and neuroinflammatory signaling, shaping neuronal resilience and plasticity. Altered Nurr1 expression or activity has been linked to dysfunction of midbrain dopaminergic circuits and changes in microglial activation states, making it a key node for studies of neurodegeneration-relevant mechanisms. As a transcriptional regulator, Nurr1 also influences catecholamine biosynthesis and dopamine handling genes, supporting research on neuronal identity and signaling homeostasis.

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

Nurr1 CRISPR Activation Plasmid (r) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the 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 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 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 expression.

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