PC-PLD1 CRISPR Activation Plasmid (h): sc-402528-ACT

Human PLD1 encodes phospholipase D1 (PC-PLD1), a membrane-associated enzyme that hydrolyzes phosphatidylcholine to generate phosphatidic acid, a lipid second messenger that regulates membrane trafficking, cytoskeletal remodeling, and vesicle secretion. PLD1-derived phosphatidic acid influences signal transduction pathways including mTOR and small GTPase networks (ARF and Rho family), thereby shaping endocytosis, exocytosis, and receptor recycling. Through its roles in platelet activation, inflammatory signaling, and growth factor–driven responses, PLD1 has been studied in contexts such as tumor cell invasion, thrombosis biology, and neuroinflammation. Altered PLD1 activity and lipid signaling dynamics are also linked to changes in autophagy and mitochondrial stress responses, making it a useful target for mechanistic studies of cellular homeostasis.

PC-PLD1 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLD1 expression without altering the underlying DNA sequence.

PC-PLD1 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLD1 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 PLD1 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PC-PLD1 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLD1 locus and enabling the study of PC-PLD1-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PC-PLD1 pathway restoration in tumor cells with silenced or reduced PLD1 expression.

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