CLN5 CRISPR Activation Plasmid (h): sc-408473-ACT

CLN5 encodes a soluble lysosomal glycoprotein implicated in maintaining lysosome homeostasis and efficient turnover of macromolecules within the endo-lysosomal system. CLN5 participates in lysosomal trafficking and proteostasis pathways that influence autophagy-lysosome function and cellular clearance, processes central to neuronal health and long-lived cell types. Disruption of CLN5 function is linked to neuronal ceroid lipofuscinosis, a neurodegenerative lysosomal storage disorder characterized by accumulation of autofluorescent storage material and progressive cellular dysfunction. As a result, CLN5 is frequently studied in models of lysosomal biology, neurodegeneration, and stress responses associated with impaired degradative capacity.

CLN5 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous CLN5 expression without altering the underlying DNA sequence.

CLN5 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the CLN5 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 CLN5 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous CLN5 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native CLN5 locus and enabling the study of CLN5-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of CLN5 pathway restoration in tumor cells with silenced or reduced CLN5 expression.

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