Real-time genome imaging of host interactions in adeno-associated virus genome release
Summary: Adeno-associated virus (AAVs) with a self-complementary genome (sc) comprising a gene of interest are used in gene therapy. Their efficiency is limited but the molecular factors contributing to this restriction are poorly understood. We utilized scAAV2 containing a fluorescent protein-bindi...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-06-01
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| Series: | iScience |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2589004225008855 |
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| author | Luisa F. Bustamante-Jaramillo Lei Yue Joshua Fingal Gustaf Rydell Maria Johansson Tomas Edreira Oliver J. Müller Susanne Hille Martin Müller Franck Gallardo Qingxin Chen Marie-Lise Blondot Michael Kann |
| author_facet | Luisa F. Bustamante-Jaramillo Lei Yue Joshua Fingal Gustaf Rydell Maria Johansson Tomas Edreira Oliver J. Müller Susanne Hille Martin Müller Franck Gallardo Qingxin Chen Marie-Lise Blondot Michael Kann |
| author_sort | Luisa F. Bustamante-Jaramillo |
| collection | DOAJ |
| description | Summary: Adeno-associated virus (AAVs) with a self-complementary genome (sc) comprising a gene of interest are used in gene therapy. Their efficiency is limited but the molecular factors contributing to this restriction are poorly understood. We utilized scAAV2 containing a fluorescent protein-binding anchor sequence on its genome allowing visualization of released genomes by time-lapse microscopy. Pairing this technique with capsid staining, we showed that scAAV2 genome release was initiated by a partial genome exposure, triggered by elevated calcium levels while the capsids interacted with proteins of the nuclear pore. Genome release occurred subsequently requiring Rad52 decamerization in the vicinity of the host chromatin. A fraction of the released genomes was degraded by Mre11, an essential factor for chromatin stability, and cellular DNA double-strand breaks. These steps were key-factors limiting transduction, suggesting that temporary modulation of DNA damage-response-proteins is a promising way to increase scAAV efficiency in therapy. |
| format | Article |
| id | doaj-art-3f986bc410a54ad4bb9467efc9c82b1c |
| institution | Kabale University |
| issn | 2589-0042 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | iScience |
| spelling | doaj-art-3f986bc410a54ad4bb9467efc9c82b1c2025-08-20T03:46:45ZengElsevieriScience2589-00422025-06-0128611262410.1016/j.isci.2025.112624Real-time genome imaging of host interactions in adeno-associated virus genome releaseLuisa F. Bustamante-Jaramillo0Lei Yue1Joshua Fingal2Gustaf Rydell3Maria Johansson4Tomas Edreira5Oliver J. Müller6Susanne Hille7Martin Müller8Franck Gallardo9Qingxin Chen10Marie-Lise Blondot11Michael Kann12Department of Infectious Diseases, Institute of Biomedicine, Gothenburg University, Gothenburg, Sweden; Corresponding authorDepartment of Infectious Diseases, Institute of Biomedicine, Gothenburg University, Gothenburg, SwedenDepartment of Infectious Diseases, Institute of Biomedicine, Gothenburg University, Gothenburg, SwedenDepartment of Infectious Diseases, Institute of Biomedicine, Gothenburg University, Gothenburg, SwedenDepartment of Infectious Diseases, Institute of Biomedicine, Gothenburg University, Gothenburg, SwedenDepartment of Chemistry and Molecular Biology, Gothenburg University, Gothenburg, SwedenDepartment of Internal Medicine V, University of Kiel, and German Centre for Cardiovascular Research, Partner Site Hamburg/Kiel/Lübeck, Germany, Kiel, GermanyDepartment of Internal Medicine V, University of Kiel, and German Centre for Cardiovascular Research, Partner Site Hamburg/Kiel/Lübeck, Germany, Kiel, GermanyInfection, Inflammation and Cancer Program, German Cancer Research Center (DKFZ), Heidelberg, GermanyNeo Vir Tech SAS, Centre Pierre Potier, Toulouse, FranceInfection, Inflammation and Cancer Program, German Cancer Research Center (DKFZ), Heidelberg, GermanyCNRS UMR 5234, Université de Bordeaux, Bordeaux, FranceDepartment of Infectious Diseases, Institute of Biomedicine, Gothenburg University, Gothenburg, Sweden; Sahlgrenska Academy, Gothenburg, Sweden; Corresponding authorSummary: Adeno-associated virus (AAVs) with a self-complementary genome (sc) comprising a gene of interest are used in gene therapy. Their efficiency is limited but the molecular factors contributing to this restriction are poorly understood. We utilized scAAV2 containing a fluorescent protein-binding anchor sequence on its genome allowing visualization of released genomes by time-lapse microscopy. Pairing this technique with capsid staining, we showed that scAAV2 genome release was initiated by a partial genome exposure, triggered by elevated calcium levels while the capsids interacted with proteins of the nuclear pore. Genome release occurred subsequently requiring Rad52 decamerization in the vicinity of the host chromatin. A fraction of the released genomes was degraded by Mre11, an essential factor for chromatin stability, and cellular DNA double-strand breaks. These steps were key-factors limiting transduction, suggesting that temporary modulation of DNA damage-response-proteins is a promising way to increase scAAV efficiency in therapy.http://www.sciencedirect.com/science/article/pii/S2589004225008855Cellular therapyGenomicsVirologyGenomic analysis |
| spellingShingle | Luisa F. Bustamante-Jaramillo Lei Yue Joshua Fingal Gustaf Rydell Maria Johansson Tomas Edreira Oliver J. Müller Susanne Hille Martin Müller Franck Gallardo Qingxin Chen Marie-Lise Blondot Michael Kann Real-time genome imaging of host interactions in adeno-associated virus genome release iScience Cellular therapy Genomics Virology Genomic analysis |
| title | Real-time genome imaging of host interactions in adeno-associated virus genome release |
| title_full | Real-time genome imaging of host interactions in adeno-associated virus genome release |
| title_fullStr | Real-time genome imaging of host interactions in adeno-associated virus genome release |
| title_full_unstemmed | Real-time genome imaging of host interactions in adeno-associated virus genome release |
| title_short | Real-time genome imaging of host interactions in adeno-associated virus genome release |
| title_sort | real time genome imaging of host interactions in adeno associated virus genome release |
| topic | Cellular therapy Genomics Virology Genomic analysis |
| url | http://www.sciencedirect.com/science/article/pii/S2589004225008855 |
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