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A robust and all-inclusive pipeline for shuffling of adeno-associated viruses

Adeno-associated viruses (AAV) are attractive templates for engineering of synthetic gene delivery vectors. A particularly powerful technology for breeding of novel vectors with improved properties is DNA family shuffling, i.e., generation of chimeric capsids by homology-driven DNA recombination. He...

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Bibliographic Details
Main Authors: Herrmann, Anne-Kathrin (Author) , Kienle, Eike (Author) , Große, Stefanie (Author) , El Andari, Jihad (Author) , Botta, Julia (Author) , Schürmann, Nina (Author) , Wiedtke, Ellen (Author) , Niopek, Dominik (Author) , Grimm, Dirk (Author)
Format: Article (Journal)
Language:English
Published: 2019
In: ACS synthetic biology
Year: 2018, Volume: 8, Issue: 1, Pages: 194-206
ISSN:2161-5063
DOI:10.1021/acssynbio.8b00373
Online Access:Verlag, Volltext: https://doi.org/10.1021/acssynbio.8b00373
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Author Notes:Anne-Kathrin Herrmann, Christian Bender, Eike Kienle, Stefanie Grosse, Jihad El Andari, Julia Botta, Nina Schürmann, Ellen Wiedtke, Dominik Niopek, and Dirk Grimm
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Summary:Adeno-associated viruses (AAV) are attractive templates for engineering of synthetic gene delivery vectors. A particularly powerful technology for breeding of novel vectors with improved properties is DNA family shuffling, i.e., generation of chimeric capsids by homology-driven DNA recombination. Here, to make AAV DNA shuffling available to a wider community, we present a robust experimental and bioinformatical pipeline comprising: (i) standardized and partially codon-optimized plasmids carrying 12 different AAV capsid genes; (ii) a scalable protocol including troubleshooting guide for viral library production; and (iii) the freely available software SALANTO for comprehensive analysis of chimeric AAV DNA and protein sequences. Moreover, we describe a set of 12 premade and ready-to-use AAV libraries. Finally, we demonstrate the usefulness of DNA barcoding technology to trace AAV capsid libraries within a complex mixture. Our protocols and resources facilitate the implementation and tailoring of AAV evolution technology in any laboratory interested in customized viral gene transfer.
Item Description:Published: December 4, 2018
Gesehen am 05.07.2019
Physical Description:Online Resource
ISSN:2161-5063
DOI:10.1021/acssynbio.8b00373

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