endobrevin CRISPR Activation Plasmid (h): sc-403438-ACT

Human VAMP8 encodes endobrevin, a vesicle-associated SNARE that mediates membrane fusion events required for endosomal trafficking, regulated exocytosis, and granule secretion. Endobrevin participates in SNARE complex assembly with plasma membrane partners to control vesicle docking and fusion in immune cells, platelets, and epithelial compartments, influencing processes such as degranulation, cytokine release, and mucin secretion. Through these roles, VAMP8 impacts vesicular transport pathways that shape inflammatory signaling, hemostasis, and host defense. Dysregulated VAMP8-dependent trafficking has been linked in the literature to altered secretory function and inflammatory phenotypes, supporting its relevance for mechanistic studies of vesicle fusion and secretion-associated disease biology.

endobrevin CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous VAMP8 expression without altering the underlying DNA sequence.

endobrevin CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the VAMP8 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 VAMP8 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous endobrevin expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native VAMP8 locus and enabling the study of endobrevin-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of endobrevin pathway restoration in tumor cells with silenced or reduced VAMP8 expression.

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