UGT1A5 CRISPR Activation Plasmid (h): sc-402838-ACT

UGT1A5 encodes a UDP-glucuronosyltransferase that catalyzes glucuronidation reactions in Phase II metabolism, increasing the solubility and promoting clearance of diverse endogenous and xenobiotic compounds. As part of the UGT1A locus, UGT1A5 contributes to cellular detoxification networks that interface with hepatic and extrahepatic metabolism, redox balance, and chemical stress responses. Variation in UGT1A family activity is widely studied for its impact on drug disposition, metabolic homeostasis, and susceptibility to toxicity from environmental exposures. Dysregulated glucuronidation capacity is also relevant to models of liver dysfunction and to cancer biology where altered conjugation can influence signaling metabolites and xenobiotic handling.

UGT1A5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UGT1A5 expression without altering the underlying DNA sequence.

UGT1A5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UGT1A5 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 UGT1A5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UGT1A5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UGT1A5 locus and enabling the study of UGT1A5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UGT1A5 pathway restoration in tumor cells with silenced or reduced UGT1A5 expression.

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