Synaptogyrin-2 CRISPR Activation Plasmid (h): sc-404687-ACT

Human SYNGR2 encodes synaptogyrin-2, a tetraspan synaptic vesicle membrane protein implicated in presynaptic organization and activity-dependent neurotransmitter release. Synaptogyrin family members contribute to vesicle biogenesis, trafficking, and recycling by influencing synaptic vesicle composition and dynamics within the synaptic transmission program. Through these roles, SYNGR2 expression and regulation are relevant to studies of neuronal excitability, synaptic plasticity, and circuit function, with potential relevance to neuropsychiatric and neurodevelopmental disease mechanisms where vesicle cycling and presynaptic signaling are perturbed. Measuring SYNGR2-dependent changes in vesicle markers, release probability, and endocytosis kinetics can help connect gene regulation to cellular phenotypes in neuronal and neurosecretory models.

Synaptogyrin-2 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SYNGR2 expression without altering the underlying DNA sequence.

Synaptogyrin-2 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SYNGR2 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 SYNGR2 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Synaptogyrin-2 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SYNGR2 locus and enabling the study of Synaptogyrin-2-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Synaptogyrin-2 pathway restoration in tumor cells with silenced or reduced SYNGR2 expression.

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