PLIC-1 CRISPR Activation Plasmid (h): sc-408872-ACT

UBQLN1 encodes ubiquilin-1 (PLIC-1), a ubiquitin-like/ubiquitin-associated adaptor that couples polyubiquitinated substrates to the 26S proteasome and helps maintain protein homeostasis. PLIC-1 participates in ubiquitin-dependent proteolysis, ER-associated degradation (ERAD), and stress-responsive proteostasis pathways, influencing turnover of misfolded or aggregation-prone proteins. Through its roles in protein quality control and trafficking, UBQLN1 is frequently studied in the context of neurodegenerative protein aggregation and cellular stress phenotypes. Altered UBQLN1 function or expression has also been linked to dysregulated proteostasis in cancer- and inflammation-relevant signaling environments, supporting its use in pathway and mechanism-focused research.

PLIC-1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UBQLN1 expression without altering the underlying DNA sequence.

PLIC-1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UBQLN1 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 UBQLN1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLIC-1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UBQLN1 locus and enabling the study of PLIC-1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLIC-1 pathway restoration in tumor cells with silenced or reduced UBQLN1 expression.

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