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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| Hauptverfasser: | , , , , , , , , |
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| Dokumenttyp: | Article (Journal) |
| Sprache: | Englisch |
| Veröffentlicht: |
2019
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| In: |
ACS synthetic biology
Year: 2018, Jahrgang: 8, Heft: 1, Pages: 194-206 |
| ISSN: | 2161-5063 |
| DOI: | 10.1021/acssynbio.8b00373 |
| Online-Zugang: | Verlag, Volltext: https://doi.org/10.1021/acssynbio.8b00373
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| Verfasserangaben: | 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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| 520 | |a 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. | ||
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