Nur77 CRISPR Activation Plasmid (h): sc-418320-ACT

NR4A1 encodes the orphan nuclear receptor Nur77, an immediate-early transcription factor that rapidly couples extracellular cues to gene expression programs controlling cell fate. Nur77 integrates signaling downstream of MAPK, PI3K/AKT, and calcium-dependent pathways to regulate apoptosis, proliferation, and metabolic adaptation, and it can modulate inflammatory transcriptional networks through crosstalk with NF-κB and AP-1. In immune cells, Nur77 is a key regulator of T cell receptor–driven activation, tolerance, and differentiation, influencing cytokine programs and stress responses. Dysregulated NR4A1 expression or activity has been associated with inflammatory conditions and cancer biology, supporting its use as a mechanistic node for pathway-focused research.

Nur77 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR4A1 expression without altering the underlying DNA sequence.

Nur77 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR4A1 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 NR4A1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Nur77 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR4A1 locus and enabling the study of Nur77-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Nur77 pathway restoration in tumor cells with silenced or reduced NR4A1 expression.

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