PSS2 CRISPR Activation Plasmid (h): sc-404133-ACT

Human PTDSS2 encodes phosphatidylserine synthase 2 (PSS2), an endoplasmic reticulum membrane enzyme that catalyzes phosphatidylserine production via base-exchange with phosphatidylethanolamine. By controlling cellular phosphatidylserine abundance, PSS2 helps regulate membrane biogenesis, organelle homeostasis, vesicular trafficking, and lipid-dependent signaling processes that influence apoptosis and phagocytic clearance. PTDSS2 activity links closely to phospholipid remodeling pathways and ER–mitochondria lipid exchange, shaping mitochondrial function and cellular stress responses. Dysregulation of phosphatidylserine metabolism has been connected to altered growth and survival phenotypes in cancer and to broader metabolic and neurobiology-relevant membrane defects, making PTDSS2 a useful target for mechanistic studies.

PSS2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous PTDSS2 expression without altering the underlying DNA sequence.

PSS2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the PTDSS2 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 PTDSS2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PSS2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native PTDSS2 locus and enabling the study of PSS2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PSS2 pathway restoration in tumor cells with silenced or reduced PTDSS2 expression.

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