pAAVdual-GFAP-CcdB

The pAAVdual-GFAP-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 GFAP promoter refers to the regulatory sequence that controls the expression of the Glial Fibrillary Acidic Protein (GFAP) gene. GFAP is a key intermediate filament protein that is predominantly expressed in astrocytes, which are star-shaped glial cells in the central nervous system (CNS). The GFAP promoter is a glial-specific regulatory element that drives gene expression primarily in astrocytes within the central nervous system. It is a powerful tool in neuroscience research, gene therapy, and the development of transgenic models for studying astrocyte function, CNS development, and neurodegenerative diseases. Its specificity to astrocytes makes it invaluable for targeting gene expression in studies focused on glial cell biology, CNS health, and pathology. 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-GFAP-CcdB destination plasmid along with the pENTR-GOI vector from the ORFeome collections, researchers can efficiently generate pAAVdual-GFAP-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