RXRα CRISPR Activation Plasmid (h): sc-400342-ACT

Retinoid X receptor alpha (RXRA; RXRα) is a ligand-activated nuclear receptor that functions as a central dimerization partner for multiple transcription factors, including PPARs, LXRs, FXR, VDR, and RARs, to regulate gene programs controlling lipid and glucose metabolism, bile acid signaling, inflammation, and cellular differentiation. RXRα modulates chromatin-dependent transcription through recruitment of coactivators and corepressors, integrating retinoid and metabolic cues across tissues. Dysregulated RXRA activity or altered RXRα signaling networks have been linked to metabolic dysfunction, aberrant immune responses, and context-dependent effects on proliferation and differentiation in cancer biology. As a transcriptional hub, RXRA is widely used to interrogate nuclear receptor crosstalk and transcriptional circuitry in hepatocytes, adipocytes, macrophages, and cancer cell models.

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

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

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