NR5A2 CRISPR Activation Plasmid (h): sc-402656-ACT

NR5A2 (also known as liver receptor homolog-1, LRH-1) is a human orphan nuclear receptor transcription factor that regulates gene programs controlling cholesterol and bile acid metabolism, steroidogenesis, and cellular differentiation. It functions through sequence-specific DNA binding and co-regulator recruitment to shape transcriptional networks involved in lipid homeostasis and epithelial tissue maintenance. NR5A2 activity intersects with metabolic signaling and developmental processes, and altered expression has been associated with dysregulated proliferation, inflammation-linked transcriptional states, and tumor biology in multiple tissue contexts. These features make NR5A2 a useful node for studying transcriptional control of metabolism, lineage commitment, and disease-relevant regulatory circuitry.

NR5A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous NR5A2 expression without altering the underlying DNA sequence.

NR5A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the NR5A2 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 NR5A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous NR5A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native NR5A2 locus and enabling the study of NR5A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of NR5A2 pathway restoration in tumor cells with silenced or reduced NR5A2 expression.

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