Peroxin 13 CRISPR Activation Plasmid (h): sc-404060-ACT

PEX13 encodes peroxin 13, an integral peroxisomal membrane component required for peroxisome biogenesis and matrix protein import. Peroxin 13 functions within the peroxisomal protein translocation machinery by coordinating peroxins that recognize PTS1/PTS2 cargo receptors, supporting import of enzymes needed for fatty-acid β-oxidation and reactive oxygen species metabolism. Disruption of PEX13 perturbs peroxisomal homeostasis, alters lipid handling and redox balance, and is associated with peroxisome biogenesis disorder phenotypes linked to impaired organelle assembly. Human PEX13 is therefore a useful target for studying peroxisomal import dynamics, metabolic rewiring, and organelle stress signaling in cell models.

Peroxin 13 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PEX13 expression without altering the underlying DNA sequence.

Peroxin 13 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PEX13 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 PEX13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Peroxin 13 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PEX13 locus and enabling the study of Peroxin 13-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Peroxin 13 pathway restoration in tumor cells with silenced or reduced PEX13 expression.

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