ATP13A2 CRISPR Activation Plasmid (h): sc-404770-ACT

ATP13A2 encodes a lysosomal P5-type ATPase implicated in maintaining endolysosomal homeostasis, including lysosomal membrane function, cation and polyamine handling, and vesicular trafficking. By supporting lysosome-dependent quality control, ATP13A2 influences autophagy–lysosome flux and proteostasis pathways that intersect with mitochondrial stress responses and oxidative damage. Disruption or dysregulation of ATP13A2 has been linked to neurodegenerative phenotypes, with recurrent associations to juvenile-onset parkinsonism and related lysosomal storage–like defects observed in cellular models. As a result, ATP13A2 is widely studied for its role in neuronal vulnerability, aggregation-prone protein clearance, and endolysosomal signaling networks.

ATP13A2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous ATP13A2 expression without altering the underlying DNA sequence.

ATP13A2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the ATP13A2 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 ATP13A2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous ATP13A2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native ATP13A2 locus and enabling the study of ATP13A2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of ATP13A2 pathway restoration in tumor cells with silenced or reduced ATP13A2 expression.

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