PC-PLD2 CRISPR Activation Plasmid (h): sc-402056-ACT

Human PLD2 encodes phospholipase D2, a membrane-associated enzyme that hydrolyzes phosphatidylcholine to generate phosphatidic acid, a lipid second messenger that shapes membrane dynamics and signaling. PC-PLD2 activity supports receptor-driven signal transduction, endocytosis and vesicle trafficking, and cytoskeletal remodeling, integrating inputs from growth factor and GPCR pathways with downstream networks such as PI3K–AKT and mTOR. Through regulation of cell migration, adhesion, and inflammatory mediator production, PLD2 contributes to processes relevant to oncogenic signaling, immune cell function, and metabolic stress responses. Altered PLD2 expression or activity has been associated with dysregulated proliferation and motility in tumor biology and with inflammatory pathophysiology, making it a useful node for mechanistic studies of lipid signaling.

PC-PLD2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PLD2 expression without altering the underlying DNA sequence.

PC-PLD2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PLD2 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 PLD2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PC-PLD2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PLD2 locus and enabling the study of PC-PLD2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PC-PLD2 pathway restoration in tumor cells with silenced or reduced PLD2 expression.

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