Synaptotagmin IV CRISPR Activation Plasmid (h): sc-403314-ACT

SYT4 encodes Synaptotagmin IV, a membrane-associated C2 domain protein that localizes to secretory vesicles and modulates Ca²⁺-dependent exocytosis in neurons and other excitable cells. Unlike canonical synaptotagmins that act as rapid Ca²⁺ sensors, Synaptotagmin IV is often linked to activity-dependent tuning of vesicle fusion probability, synaptic plasticity, and regulated release of neuropeptides and neurotransmitters. SYT4 function intersects with SNARE-mediated membrane fusion, vesicle trafficking, and stimulus-secretion coupling pathways that influence neuronal network adaptation. Altered expression or regulation of vesicle release machinery, including SYT4, has been associated with neurodevelopmental and neuropsychiatric phenotypes and is studied in the context of synaptic dysfunction.

Synaptotagmin IV CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SYT4 expression without altering the underlying DNA sequence.

Synaptotagmin IV CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SYT4 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 SYT4 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Synaptotagmin IV expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SYT4 locus and enabling the study of Synaptotagmin IV-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Synaptotagmin IV pathway restoration in tumor cells with silenced or reduced SYT4 expression.

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