Key result
Protocol outlines serotype selection and delivery methods for recombinant adeno-associated virus vectors to murine lungs.
Why the study?
Efficient delivery of recombinant AAV vectors to the lung requires understanding of serotype selection and delivery methods for targeted gene transfer.
Facilitates reproducible AAV gene transfer in murine lung models; extends preclinical tools for cardiovascular researchers while leaving clinical translation open.
Adeno-associated virus (AAV) is a small replication-deficient DNA virus belonging to the Parvovirinae family. It has a single-stranded ∼4.7-kb genome. Recombinant AAV (rAAV) is created by replacing the viral rep and cap genes with the transgene of interest along with promoter and polyadenylation sequences. The short viral inverted terminal repeats must remain intact for replication and packaging in production, as well as vector genome processing and persistence in the transduction process. The AAV capsid (serotype) determines the tissue tropism of the rAAV vector. In this unit we will discuss serotype selection for lung targeting along with the factors effecting efficient delivery of rAAV vectors to the murine lung. Detailed procedures for lung delivery (intranasal, orotracheal, and surgical tracheal injection), sample collection, and post-mortem tissue processing will be described.
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Gruntman et al. (2012) reported a review. Recombinant Adeno-Associated Virus (rAAV) gene transfer was evaluated. This protocol describes serotype selection and detailed procedures for delivering recombinant adeno-associated virus vectors to the murine lung.
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