USPL1 CRISPR Activation Plasmid (h2): sc-408809-ACT-2

Human USPL1 (ubiquitin-specific peptidase-like 1) encodes a SUMO isopeptidase that localizes predominantly to Cajal bodies and regulates SUMO-dependent protein turnover and nuclear organization. USPL1 supports spliceosomal snRNP biogenesis and RNA processing by controlling SUMOylation dynamics of nuclear factors, linking it to transcriptional regulation and cell-cycle–coupled genome maintenance programs. Dysregulated SUMO homeostasis and altered USPL1 activity have been associated with proliferative phenotypes and cancer-relevant pathways, reflecting the sensitivity of nuclear RNA-processing hubs to SUMO protease function. Targeted editing of USPL1 enables mechanistic studies of SUMO pathway circuitry, Cajal body integrity, and context-specific effects on splicing, gene expression, and cellular fitness in human model systems.

USPL1 CRISPR Activation Plasmid (h2) provides a targeted, non-destructive approach to upregulating endogenous USPL1 expression without altering the underlying DNA sequence.

USPL1 CRISPR Activation Plasmid (h2) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the USPL1 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 USPL1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous USPL1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native USPL1 locus and enabling the study of USPL1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of USPL1 pathway restoration in tumor cells with silenced or reduced USPL1 expression.

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