CYP2R1 CRISPR Activation Plasmid (h): sc-404463-ACT
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- CYP2R1 CRISPR Activation Plasmid (h) is a synergistic activation mediator (SAM) transcription activation system designed to specifically upregulate gene expression
- CYP2R1 CRISPR Activation Plasmid (h) consists of three plasmids at a 1:1:1 mass ratio: a plasmid encoding the deactivated Cas9 (dCas9) nuclease (D10A and N863A) fused to the transactivation domain VP64, and a blasticidin resistance gene; a plasmid encoding the MS2-p65-HSF1 fusion protein, and a hygromycin resistance gene; a plasmid encoding a target-specific 20 nt guide RNA fused to two MS2 RNA aptamers, and a puromycin resistance gene
- The resulting SAM complex binds to a site-specific region approximately 200-250 nt upstream of the transcriptional start site and provides robust recruitment of transcription factors for highly efficient gene activation
- gRNAs encoded by CYP2R1 CRISPR Activation Plasmid (h) and CYP2R1 CRISPR Activation Plasmid (h2) target distinct regulatory regions upstream of the CYP2R1 transcriptional start site. One or both designs may be available
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
CYP2R1 CRISPR Activation Plasmid (h) | sc-404463-ACT | 20 µg | $397.00 |
Human CYP2R1 encodes a microsomal cytochrome P450 monooxygenase that functions as a principal 25-hydroxylase in vitamin D metabolism, converting cholecalciferol and ergocalciferol to 25-hydroxyvitamin D in the endoplasmic reticulum. This enzymatic step is central to steroid and lipid oxidation pathways and contributes to systemic regulation of calcium-phosphate homeostasis through downstream vitamin D endocrine signaling. Variation or reduced activity of CYP2R1 has been linked to low circulating 25-hydroxyvitamin D levels and related phenotypes affecting bone and mineral metabolism. CYP2R1 is therefore relevant for mechanistic studies of hepatic vitamin D biotransformation, cytochrome P450 redox biology, and nutrient-hormone signaling networks.
CYP2R1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CYP2R1 expression without altering the underlying DNA sequence.
CYP2R1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CYP2R1 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 CYP2R1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CYP2R1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CYP2R1 locus and enabling the study of CYP2R1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CYP2R1 pathway restoration in tumor cells with silenced or reduced CYP2R1 expression.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.