Rebecca Christine Feiner, Julian Teschner, Kathrin E. Teschner, Marco T. Radukic, Tobias Baumann, Sven Hagen, Yvonne Hannappel, Niklas Biere, Dario Anselmetti, Katja Maren Arndt, Kristian Mark Müller
- Recombinant adeno-associated viruses (rAAV) provide outstanding options for customization and superior capabilities for gene therapy. To access their full potential, facile genetic manipulation is pivotal, including capsid loop modifications. Therefore, we assessed capsid tolerance to modifications of the structural VP proteins in terms of stability and plasticity. Flexible glycine-serine linkers of increasing sizes were, at the genetic level, introduced into the 587 loop region of the VP proteins of serotype 2, the best studied AAV representative. Analyses of biological function and thermal stability with respect to genome release of viral particles revealed structural plasticity. In addition, insertion of the 29 kDa enzyme beta-lactamase into the loop region was tested with a complete or a mosaic modification setting. For the mosaic approach, investigation of VP2 trans expression revealed that a Kozak sequence was required to prevent leaky scanning. Surprisingly, even the full capsid modification with beta-lactamase allowed for theRecombinant adeno-associated viruses (rAAV) provide outstanding options for customization and superior capabilities for gene therapy. To access their full potential, facile genetic manipulation is pivotal, including capsid loop modifications. Therefore, we assessed capsid tolerance to modifications of the structural VP proteins in terms of stability and plasticity. Flexible glycine-serine linkers of increasing sizes were, at the genetic level, introduced into the 587 loop region of the VP proteins of serotype 2, the best studied AAV representative. Analyses of biological function and thermal stability with respect to genome release of viral particles revealed structural plasticity. In addition, insertion of the 29 kDa enzyme beta-lactamase into the loop region was tested with a complete or a mosaic modification setting. For the mosaic approach, investigation of VP2 trans expression revealed that a Kozak sequence was required to prevent leaky scanning. Surprisingly, even the full capsid modification with beta-lactamase allowed for the assembly of capsids with a concomitant increase in size. Enzyme activity assays revealed lactamase functionality for both rAAV variants, which demonstrates the structural robustness of this platform technology.…
MetadatenVerfasserangaben: | Rebecca Christine FeinerORCiDGND, Julian TeschnerORCiDGND, Kathrin E. TeschnerGND, Marco T. Radukic, Tobias BaumannGND, Sven HagenORCiD, Yvonne HannappelORCiD, Niklas BiereORCiD, Dario AnselmettiORCiDGND, Katja Maren ArndtORCiDGND, Kristian Mark MüllerGND |
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DOI: | https://doi.org/10.3390/ijms20225702 |
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ISSN: | 1422-0067 |
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Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/31739438 |
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Titel des übergeordneten Werks (Englisch): | International journal of molecular sciences |
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Verlag: | MDPI |
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Verlagsort: | Basel |
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Publikationstyp: | Wissenschaftlicher Artikel |
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Sprache: | Englisch |
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Jahr der Erstveröffentlichung: | 2019 |
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Erscheinungsjahr: | 2019 |
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Datum der Freischaltung: | 11.10.2020 |
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Freies Schlagwort / Tag: | adeno-associated-virus; beta-lactamase; capsid stability; inverted terminal repeat (ITR); loop modification |
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Band: | 20 |
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Ausgabe: | 22 |
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Seitenanzahl: | 19 |
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Fördernde Institution: | German Research Foundation (DFG)German Research Foundation (DFG); Bielefeld University |
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Organisationseinheiten: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
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DDC-Klassifikation: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
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Peer Review: | Referiert |
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Publikationsweg: | Open Access |
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| Open Access / Gold Open-Access |
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| DOAJ gelistet |
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