Rab 3A CRISPR Activation Plasmid (h): sc-402633-ACT

RAB3A encodes the small GTPase Rab 3A, a synaptic vesicle–associated regulator of Ca²⁺-dependent exocytosis that coordinates vesicle docking, priming, and fusion at presynaptic terminals. Cycling between GDP- and GTP-bound states, Rab 3A integrates with Rab effector networks and SNARE-associated machinery to control neurotransmitter release and activity-dependent vesicle trafficking. These processes link RAB3A to neuronal signaling, synaptic plasticity, and regulated secretion pathways in neuroendocrine contexts. Dysregulation of vesicle trafficking and synaptic release dynamics is relevant to neurodevelopmental and neurodegenerative disease mechanisms, supporting RAB3A as a target for studying presynaptic function.

Rab 3A CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous RAB3A expression without altering the underlying DNA sequence.

Rab 3A CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the RAB3A 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 RAB3A transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous Rab 3A expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native RAB3A locus and enabling the study of Rab 3A-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of Rab 3A pathway restoration in tumor cells with silenced or reduced RAB3A expression.

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