Abstract
Recombinant adeno-associated virus (rAAV) has been widely used for gene therapy. AAV-mediated gene transfer leads to durable protein expression in non-proliferating targeted tissues, which enables long-term modulation of gene expression. Here we describe a rAAV production protocol based on PEI-mediated triple transfection of HEK293T cells, followed by purification by iodixanol density gradient ultracentrifugation. Viral yield varies, depending on the size of the viral genome, but, typically, a yield of 3E11 viral genome (vg) can be achieved using the described protocol. Our results showed that injection of rAAV9 significantly transduces cardiac cells, which supports rAAV9 being an effective tool for gene delivery in the heart in vivo.
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Acknowledgments
WTP was supported by funding from the Barth Syndrome Foundation and NIH (HL146634 and HL128694).
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Wang, S., Guo, Y., Pu, W.T. (2021). AAV Gene Transfer to the Heart. In: Poss, K.D., Kühn, B. (eds) Cardiac Regeneration. Methods in Molecular Biology, vol 2158. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0668-1_20
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DOI: https://doi.org/10.1007/978-1-0716-0668-1_20
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