USP6 CRISPR Activation Plasmid (h): sc-406687-ACT

USP6 (ubiquitin specific peptidase 6) encodes a deubiquitinating enzyme that modulates protein stability and signaling output by reversing ubiquitin conjugation. Through regulation of ubiquitin-dependent turnover and trafficking, USP6 can influence pathways linked to receptor signaling, cytoskeletal remodeling, and inflammatory transcriptional programs, including NF-κB–associated responses. Dysregulated USP6 expression is strongly associated with benign mesenchymal neoplasms such as aneurysmal bone cyst and nodular fasciitis, where promoter swapping events drive aberrant transcription. As a result, USP6 is widely studied for how altered deubiquitination reshapes cellular signaling networks and tumor microenvironment–relevant phenotypes.

USP6 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous USP6 expression without altering the underlying DNA sequence.

USP6 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the USP6 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 USP6 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous USP6 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native USP6 locus and enabling the study of USP6-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of USP6 pathway restoration in tumor cells with silenced or reduced USP6 expression.

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