SYP/Synaptophysin CRISPR Activation Plasmid (h): sc-400487-ACT

SYP encodes synaptophysin, an abundant integral membrane glycoprotein of synaptic vesicles that supports vesicle biogenesis, trafficking, and neurotransmitter release through regulated exocytosis. It participates in presynaptic vesicle cycling processes including endocytosis, vesicle docking, and fusion, and is commonly used as a marker of synaptic density and neuronal differentiation. Synaptophysin-linked pathways intersect with calcium-dependent synaptic transmission and activity-dependent remodeling of neuronal circuits. Altered SYP expression has been reported in neurodegenerative and neurodevelopmental disorders and is frequently evaluated in neuropathology studies as a readout of synaptic integrity.

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

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

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