UGT1A CRISPR Activation Plasmid (h): sc-417594-ACT

UGT1A8 encodes a human UDP-glucuronosyltransferase of the UGT1A family that catalyzes phase II glucuronidation of diverse xenobiotics and endogenous metabolites, increasing their aqueous solubility for biliary and renal elimination. This enzymatic activity links UGT1A8 to hepatic and extrahepatic detoxification networks, intersecting with drug metabolism pathways and cellular responses to chemical stress. Variation or altered expression within UGT1A isoforms can shift glucuronidation capacity and has been associated with inter-individual differences in metabolite clearance and susceptibility to toxicant burden. UGT1A8 is therefore relevant for mechanistic studies of conjugation biochemistry, regulation of metabolic enzymes, and context-dependent effects on cellular homeostasis.

UGT1A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UGT1A8 expression without altering the underlying DNA sequence.

UGT1A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UGT1A8 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 UGT1A8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UGT1A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UGT1A8 locus and enabling the study of UGT1A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UGT1A pathway restoration in tumor cells with silenced or reduced UGT1A8 expression.

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