PISD CRISPR Activation Plasmid (h): sc-404398-ACT

Phosphatidylserine decarboxylase (PISD) is a mitochondrial inner membrane enzyme that converts phosphatidylserine to phosphatidylethanolamine, a major phospholipid required for membrane curvature, mitochondrial dynamics, and autophagy-related membrane remodeling. By supporting phospholipid homeostasis and oxidative phosphorylation–linked mitochondrial function, PISD contributes to cellular bioenergetics and stress adaptation. Altered phosphatidylethanolamine availability has been associated with mitochondrial dysfunction, impaired mitophagy, and broader metabolic and neurodevelopmental phenotypes, making PISD regulation relevant to studies of organelle quality control. PISD is therefore frequently examined in pathways connecting lipid metabolism, membrane trafficking, and mitochondrial maintenance.

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

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

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