UGT2B CRISPR Activation Plasmid (h): sc-404208-ACT

Human UGT2B4 encodes a UDP-glucuronosyltransferase of the UGT2B subfamily that catalyzes glucuronidation of endogenous steroids, bile acid–related metabolites, and diverse xenobiotics, increasing their solubility for elimination. This enzymatic activity supports phase II metabolism in endoplasmic reticulum–associated detoxification networks and interfaces with hepatic clearance pathways regulated by nuclear receptor signaling and redox homeostasis. Variation in UGT2B4 expression or activity can shift steroid hormone disposition and xenobiotic handling, making it relevant to studies of interindividual drug metabolism, endocrine regulation, and metabolic stress phenotypes. As part of the broader UGT2B functional landscape, UGT2B4 is frequently examined alongside transport and conjugation systems that together shape cellular exposure to bioactive small molecules.

UGT2B4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UGT2B4 expression without altering the underlying DNA sequence.

UGT2B4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UGT2B4 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 UGT2B4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UGT2B4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UGT2B4 locus and enabling the study of UGT2B4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UGT2B4 pathway restoration in tumor cells with silenced or reduced UGT2B4 expression.

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