UGT1A4 CRISPR Activation Plasmid (h): sc-402659-ACT

UGT1A4 encodes a UDP-glucuronosyltransferase that catalyzes phase II glucuronidation of diverse xenobiotics and endogenous metabolites, increasing their solubility for biliary and renal elimination. As a microsomal enzyme predominantly expressed in liver, UGT1A4 contributes to cellular detoxification and metabolic homeostasis by channeling substrates into glucuronide conjugation pathways downstream of cytochrome P450 oxidation. Variation in UGT1A4 expression or activity has been associated with interindividual differences in drug metabolism and chemical sensitivity, making it relevant to pharmacogenomics and toxicology research. Experimental modulation of UGT1A4 also informs studies of hepatic differentiation, endoplasmic reticulum–linked metabolism, and adaptive responses to chemical stress.

UGT1A4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UGT1A4 expression without altering the underlying DNA sequence.

UGT1A4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UGT1A4 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 UGT1A4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UGT1A4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UGT1A4 locus and enabling the study of UGT1A4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UGT1A4 pathway restoration in tumor cells with silenced or reduced UGT1A4 expression.

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