LXR alpha/NR1H3 CRISPR Activation Plasmid (m): sc-423616-ACT

Nr1h3 encodes liver X receptor alpha (LXRα/NR1H3), a ligand-activated nuclear receptor that forms heterodimers with RXR to regulate transcriptional programs controlling cholesterol efflux, bile acid metabolism, and fatty acid synthesis. In macrophages and hepatocytes, LXRα coordinates lipid handling through targets such as ABCA1/ABCG1 and interfaces with innate immune signaling to modulate inflammatory gene expression. These pathways connect NR1H3 activity to foam cell biology, atherogenic lipid accumulation, and metabolic inflammation, making it a widely used node for studying lipid homeostasis in cardiovascular and metabolic disease models. In mouse systems, Nr1h3 perturbation is also leveraged to dissect cross-talk between sterol sensing, hepatic lipogenesis, and immune cell polarization.

LXR alpha/NR1H3 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Nr1h3 expression without altering the underlying DNA sequence.

LXR alpha/NR1H3 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Nr1h3 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 Nr1h3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LXR alpha/NR1H3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Nr1h3 locus and enabling the study of LXR alpha/NR1H3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LXR alpha/NR1H3 pathway restoration in tumor cells with silenced or reduced Nr1h3 expression.

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