UGT1A9 CRISPR Activation Plasmid (h): sc-417693-ACT

UGT1A9 encodes a human UDP-glucuronosyltransferase that catalyzes glucuronidation of diverse xenobiotics and endogenous small molecules, increasing their aqueous solubility and facilitating clearance. As a phase II metabolism enzyme, UGT1A9 contributes to hepatic and extrahepatic detoxification networks that interface with CYP-dependent oxidation, redox balance, and transporter-mediated disposition pathways. Variation in UGT1A9 expression or activity can shift cellular handling of drugs and metabolites, influencing susceptibility to metabolite-driven stress responses. In research settings, UGT1A9 serves as a key node for interrogating glucuronidation capacity, chemical biology, and inter-individual variability in biotransformation.

UGT1A9 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UGT1A9 expression without altering the underlying DNA sequence.

UGT1A9 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UGT1A9 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 UGT1A9 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UGT1A9 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UGT1A9 locus and enabling the study of UGT1A9-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UGT1A9 pathway restoration in tumor cells with silenced or reduced UGT1A9 expression.

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