pAAVdual-CaMKIIα-CcdB

The pAAVdual-CaMKIIα-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 CaMKIIα promoter refers to the regulatory sequence that controls the expression of the Calcium/Calmodulin-Dependent Protein Kinase II Alpha (CaMKIIα) gene. CaMKIIα is an essential enzyme predominantly expressed in the brain, especially in excitatory neurons of the forebrain, such as those found in the hippocampus and cortex. The CaMKIIα promoter is a neuron-specific regulatory element that drives gene expression in excitatory neurons of the forebrain, particularly in regions like the hippocampus and cortex. It is a critical tool in neuroscience research, transgenic model development, optogenetics, and potentially gene therapy. Its specificity to neurons involved in synaptic plasticity, learning, and memory makes it invaluable for studying the molecular and cellular mechanisms underlying these key brain functions. 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-CaMKIIα-CcdB destination plasmid along with the pENTR-GOI vector from the ORFeome collections, researchers can efficiently generate pAAVdual-CaMKIIα-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