Vitamin D Receptor/VDR CRISPR Activation Plasmid (m): sc-423664-ACT

Mouse Vdr encodes the vitamin D receptor (VDR), a ligand-activated nuclear receptor that heterodimerizes with RXR to bind vitamin D response elements and regulate transcriptional programs controlling calcium and phosphate homeostasis, epithelial differentiation, and immune modulation. VDR integrates signals from steroid hormone signaling with chromatin remodeling to influence cell-cycle control, barrier function, and inflammatory gene expression across multiple tissues. In biomedical research, altered Vdr activity is commonly studied in contexts of bone and mineral metabolism, intestinal physiology, and immune-mediated inflammation, where shifts in VDR-dependent transcription can reshape cytokine networks and metabolic pathways. These functions make Vdr a central node for dissecting transcriptional regulation and signaling crosstalk in development and disease-relevant phenotypes in mouse models and cultured cells.

Vitamin D Receptor/VDR CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Vdr expression without altering the underlying DNA sequence.

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

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