PRELI CRISPR Activation Plasmid (h): sc-411975-ACT

Human PRELID1 encodes PRELI, a mitochondrial intermembrane space protein implicated in maintaining mitochondrial phospholipid homeostasis and inner membrane integrity. PRELI functions within PRELI-like lipid transfer pathways that support cardiolipin-related membrane organization, oxidative phosphorylation efficiency, and mitochondrial dynamics that influence apoptosis and stress adaptation. By coupling lipid handling to respiratory chain performance, PRELI contributes to cellular energy metabolism and redox balance. Altered mitochondrial lipid composition and bioenergetic stress signaling associated with PRELID1 dysregulation are relevant to mechanistic studies of mitochondrial dysfunction observed across cancer biology and neurodegeneration-associated phenotypes.

PRELI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PRELID1 expression without altering the underlying DNA sequence.

PRELI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PRELID1 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 PRELID1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PRELI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PRELID1 locus and enabling the study of PRELI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PRELI pathway restoration in tumor cells with silenced or reduced PRELID1 expression.

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