Synaptotagmin XI CRISPR Activation Plasmid (h): sc-404323-ACT

SYT11 encodes human Synaptotagmin XI, a member of the synaptotagmin family implicated in regulation of intracellular membrane trafficking and vesicle dynamics. Synaptotagmin XI has been linked to control of exocytosis and endocytic recycling, influencing synaptic vesicle turnover and broader secretory pathway processes in neurons and other secretory cell types. Through effects on vesicle docking, fusion competence, and organelle homeostasis, SYT11 can intersect with pathways governing neurotransmission, autophagy-lysosome function, and cellular stress responses. Altered SYT11 expression or genetic variation has been associated with neurodegenerative and neuroinflammatory research contexts, supporting its use in studies of neuronal vulnerability and synaptic dysfunction mechanisms.

Synaptotagmin XI CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous SYT11 expression without altering the underlying DNA sequence.

Synaptotagmin XI CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the SYT11 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 SYT11 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Synaptotagmin XI expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native SYT11 locus and enabling the study of Synaptotagmin XI-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Synaptotagmin XI pathway restoration in tumor cells with silenced or reduced SYT11 expression.

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