Peroxin 14 CRISPR Activation Plasmid (h): sc-405075-ACT

PEX14 encodes peroxin 14, an essential component of the peroxisomal import machinery that anchors the PEX5/PEX7 receptor complexes at the peroxisome membrane to enable translocation of matrix enzymes. Through its role in peroxisome biogenesis and protein import, PEX14 supports fatty acid β-oxidation, ether lipid synthesis, and reactive oxygen species detoxification, linking it to cellular redox balance and lipid homeostasis. Altered peroxisomal import and biogenesis are associated with peroxisome biogenesis disorders and broader metabolic and neurodevelopmental phenotypes, making PEX14 regulation relevant for mechanistic studies of organelle dysfunction. In human cell models, modulation of PEX14 provides a tractable entry point for investigating peroxisome-dependent pathways and their interaction with mitochondrial and inflammatory signaling.

Peroxin 14 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PEX14 expression without altering the underlying DNA sequence.

Peroxin 14 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PEX14 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 PEX14 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Peroxin 14 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PEX14 locus and enabling the study of Peroxin 14-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Peroxin 14 pathway restoration in tumor cells with silenced or reduced PEX14 expression.

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