LAMP-2 CRISPR Activation Plasmid (m): sc-421383-ACT

Mouse Lamp2 encodes lysosome-associated membrane protein 2 (LAMP-2), a heavily glycosylated type I membrane protein enriched on the lysosomal limiting membrane that supports lysosome integrity and substrate turnover. LAMP-2 participates in autophagy–lysosome pathways, including autophagosome–lysosome fusion and chaperone-mediated autophagy, thereby influencing organelle quality control, proteostasis, and cellular stress responses. Altered LAMP-2 function is linked to lysosomal storage phenotypes and impaired autophagic flux, processes implicated in cardiomyopathy, myopathy, and neurodegeneration models. As a central component of lysosomal homeostasis, Lamp2 is frequently studied in contexts such as endolysosomal trafficking, mitochondrial quality control, and immune cell antigen processing.

LAMP-2 CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Lamp2 expression without altering the underlying DNA sequence.

LAMP-2 CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Lamp2 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 Lamp2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous LAMP-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Lamp2 locus and enabling the study of LAMP-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of LAMP-2 pathway restoration in tumor cells with silenced or reduced Lamp2 expression.

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