pAAVdual-hSyn-CcdB

The pAAVdual-hSyn-CcdB plasmid serves as a destination vector for efficient ORF cloning into the pAAVdual system using Gateway technology. This system is part of AAVdual platform, which integrates the AAV vector plasmid (pAAVtri) and the Ad helper plasmid (mini-pHelper) into a single vector. The hSyn promoter, also known as the human Synapsin 1 promoter, is a regulatory sequence used to drive gene expression specifically in neurons. Synapsin 1 is a neuron-specific phosphoprotein that plays a crucial role in the regulation of neurotransmitter release at synapses. The hSyn promoter is a neuron-specific regulatory element that drives strong and selective gene expression in neurons. It is widely used in neuroscience research, gene therapy, the development of transgenic models, and optogenetic studies. Its specificity to neurons makes it invaluable for targeting gene expression in the nervous system, facilitating research into neuronal function, connectivity, and neurological disorders. The CcdB gene in the plasmid encodes a toxic protein that disrupts DNA gyrase, a bacterial enzyme essential for DNA replication, leading to cell death. This ensures that only bacterial cells with successful recombination events—those replacing the CcdB gene—survive, facilitating positive selection during cloning. By using the pAAVdual-hSyn-CcdB destination plasmid along with the pENTR-GOI vector from the ORFeome collections, researchers can efficiently generate pAAVdual-hSyn-GOI plasmids. These plasmids can then be used for AAV packaging within the AAVdual system, enabling the production of corresponding AAV vectors.

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Background

The ORFeome collections provide a comprehensive set of open reading frames (ORFs) from a genome, with each entry vector carrying an ORF flanked by attL1 and attL2 sites. In Gateway cloning, the entry clone undergoes an LR reaction with the destination vector, where the attL and attR sites recombine. This process transfers the gene of interest from the entry vector into the destination vector, replacing the CcdB gene with the ORF, enabling successful cloning for AAV production.NameSKUPriceBuySelect all