LIMP II CRISPR Activation Plasmid (h): sc-402532-ACT

SCARB2 encodes lysosomal integral membrane protein 2 (LIMP II), a type III membrane glycoprotein enriched on late endosomes and lysosomes that supports organelle maturation, membrane trafficking, and lysosomal homeostasis. LIMP II is required for proper delivery and function of lysosomal hydrolases, including its well-characterized role as a receptor for glucocerebrosidase transport to lysosomes. Through these activities, SCARB2 influences endolysosomal dynamics, autophagy–lysosome pathway flux, and cellular responses to proteostatic and lipid metabolic stress. Dysregulation of SCARB2/LIMP II is therefore relevant to studies of lysosomal storage phenotypes, neurodegeneration-associated pathways, and host–pathogen interactions that depend on endocytic entry and lysosomal function.

LIMP II CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SCARB2 expression without altering the underlying DNA sequence.

LIMP II CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SCARB2 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 SCARB2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LIMP II expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SCARB2 locus and enabling the study of LIMP II-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LIMP II pathway restoration in tumor cells with silenced or reduced SCARB2 expression.

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