USP20 CRISPR Activation Plasmid (h): sc-406849-ACT

Human USP20 encodes a deubiquitinating enzyme (ubiquitin-specific protease 20) that removes ubiquitin chains from substrate proteins to modulate their stability, localization, and signaling output. Through regulation of ubiquitin-dependent turnover, USP20 influences processes including receptor trafficking, inflammatory signal transduction, and stress-response pathways that shape cell growth and survival. Altered deubiquitination dynamics involving USP20 have been linked in the literature to dysregulated signaling networks relevant to oncogenic transformation, immune regulation, and metabolic dysfunction, making it a useful node for mechanistic studies of proteostasis. Profiling USP20 function can therefore help clarify how ubiquitin editing tunes pathway amplitude and duration in human cells.

USP20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous USP20 expression without altering the underlying DNA sequence.

USP20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the USP20 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 USP20 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous USP20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native USP20 locus and enabling the study of USP20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of USP20 pathway restoration in tumor cells with silenced or reduced USP20 expression.

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