UPIb CRISPR Activation Plasmid (h): sc-404274-ACT

UPK1B encodes uroplakin-1B (UPIb), a tetraspanin-like integral membrane protein that assembles with other uroplakins to form urothelial plaques on the apical surface of umbrella cells. These plaques contribute to epithelial barrier integrity, membrane organization, and vesicular trafficking events that regulate surface remodeling in the urinary tract. UPK1B expression is highly enriched in differentiated urothelium and is commonly used as a marker of urothelial lineage and epithelial differentiation programs. Altered uroplakin expression patterns have been reported in urothelial pathology and are studied in the context of bladder epithelial homeostasis, metaplasia, and tumor-associated changes in differentiation state.

UPIb CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UPK1B expression without altering the underlying DNA sequence.

UPIb CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UPK1B 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 UPK1B transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous UPIb expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UPK1B locus and enabling the study of UPIb-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of UPIb pathway restoration in tumor cells with silenced or reduced UPK1B expression.

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