WDR20 CRISPR Activation Plasmid (h): sc-418266-ACT

WDR20 encodes a WD-repeat–containing regulatory protein that functions as an accessory factor for deubiquitinating enzyme complexes, helping tune ubiquitin-dependent control of protein stability and signaling. By modulating deubiquitination activity, WDR20 can influence proteostasis, receptor and kinase pathway duration, and downstream transcriptional programs that shape cell growth and stress responses. Dysregulated ubiquitin signaling and deubiquitinase regulation are broadly linked to oncogenic signaling, neurobiology, and immune homeostasis, making WDR20 a useful node for studying pathway rewiring in human cell models. Investigating WDR20 supports mechanistic work on ubiquitin dynamics, network-level pathway feedback, and context-specific regulation of signaling amplitude.

WDR20 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous WDR20 expression without altering the underlying DNA sequence.

WDR20 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the WDR20 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 WDR20 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous WDR20 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native WDR20 locus and enabling the study of WDR20-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of WDR20 pathway restoration in tumor cells with silenced or reduced WDR20 expression.

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