PLIC-2 CRISPR Activation Plasmid (h): sc-404783-ACT

Human UBQLN2 encodes the ubiquitin-like protein PLIC-2, a ubiquitin receptor and adaptor that couples polyubiquitinated substrates to the 26S proteasome to maintain proteostasis. PLIC-2 participates in ubiquitin-dependent protein quality control, ER-associated degradation, and turnover of misfolded or aggregation-prone proteins, linking it to cellular stress responses and autophagy–proteasome crosstalk. Dysregulated UBQLN2 function has been associated with aberrant protein aggregation and impaired clearance pathways relevant to neurodegenerative disease mechanisms, including ALS and frontotemporal dementia spectrum pathology. As a node in ubiquitin–proteasome signaling, UBQLN2 is frequently studied for its roles in neuronal homeostasis, cytoplasmic inclusion formation, and modulation of proteotoxic stress pathways.

PLIC-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous UBQLN2 expression without altering the underlying DNA sequence.

PLIC-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the UBQLN2 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 UBQLN2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PLIC-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native UBQLN2 locus and enabling the study of PLIC-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PLIC-2 pathway restoration in tumor cells with silenced or reduced UBQLN2 expression.

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