6533b86ffe1ef96bd12ce8f9
RESEARCH PRODUCT
A Robust and All-Inclusive Pipeline for Shuffling of Adeno-Associated Viruses.
Ellen WiedtkeEike KienleNina SchürmannAnne-kathrin HerrmannDominik NiopekChristian BenderJulia BottaJulia BottaStefanie GrosseDirk GrimmJihad El Andarisubject
0106 biological sciencesComputer sciencevirusesGenetic VectorsBiomedical EngineeringComputational biologymedicine.disease_cause01 natural sciencesBiochemistry Genetics and Molecular Biology (miscellaneous)law.inventionEvolution Molecular03 medical and health scienceschemistry.chemical_compoundPlasmidlaw010608 biotechnologymedicineGeneAdeno-associated virus030304 developmental biology0303 health sciencesShufflingGene Transfer TechniquesGeneral MedicineDependovirusDNA shufflingchemistryCapsidRecombinant DNADNAdescription
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.
| year | journal | country | edition | language |
|---|---|---|---|---|
| 2018-12-05 | ACS synthetic biology |