NOR-1 CRISPR Activation Plasmid (h): sc-401813-ACT

NR4A3 encodes NOR-1, an orphan nuclear receptor that functions as an immediate-early transcription factor integrating mitogenic and stress signals into context-dependent gene programs. NOR-1 influences cellular differentiation, proliferation, apoptosis, and metabolic adaptation by modulating transcriptional networks downstream of pathways such as MAPK/ERK and cAMP/PKA, with crosstalk to inflammatory signaling. In immune and vascular cells, NR4A3 contributes to regulation of cytokine-responsive gene expression and phenotypic switching, supporting studies of inflammation, angiogenesis, and tissue remodeling. Altered NR4A3 activity and transcriptional rewiring have been associated with oncogenic and hematologic processes, making it a useful node for investigating disease-relevant gene regulatory circuits.

NOR-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR4A3 expression without altering the underlying DNA sequence.

NOR-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR4A3 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 NR4A3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NOR-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR4A3 locus and enabling the study of NOR-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NOR-1 pathway restoration in tumor cells with silenced or reduced NR4A3 expression.

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