Synaptogyrin-3 CRISPR Activation Plasmid (h): sc-403955-ACT

SYNGR3 encodes Synaptogyrin-3, a synaptic vesicle membrane protein enriched in neurons and implicated in regulating vesicle trafficking, vesicle pool dynamics, and neurotransmitter release. Synaptogyrin family proteins participate in presynaptic membrane organization and can influence synaptic plasticity by modulating vesicle recycling and coupling to exo- and endocytic processes. Altered synaptic vesicle function and presynaptic signaling are recurrent features across neuropsychiatric and neurodegenerative disease biology, making SYNGR3 a relevant target for studying mechanisms of synaptic dysfunction. As a neuronally expressed vesicle-associated factor, SYNGR3 is useful for dissecting pathways that govern presynaptic homeostasis and activity-dependent synaptic remodeling.

Synaptogyrin-3 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SYNGR3 expression without altering the underlying DNA sequence.

Synaptogyrin-3 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SYNGR3 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 SYNGR3 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Synaptogyrin-3 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SYNGR3 locus and enabling the study of Synaptogyrin-3-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Synaptogyrin-3 pathway restoration in tumor cells with silenced or reduced SYNGR3 expression.

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