Real-time genome imaging of host interactions in adeno-associated virus genome release

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.

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Publication Details

Journal
iScience
Published
2025-05-09
DOI
https://doi.org/10.1016/j.isci.2025.112624
Citations
2
Primary Topic
Virus-based gene therapy research
Type
article
Field-Weighted Citation Impact
3.63

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article

Real-time genome imaging of host interactions in adeno-associated virus genome release

Joshua Fingal, Michael Kann, Susanne S. Hille, Martin Müller et al.
2 citations
iScience
Virus-based gene therapy research
3.63
article

Real-time genome imaging of host interactions in adeno-associated virus genome release

Joshua Fingal, Michael Kann, Susanne S. Hille, Martin Müller, Lei Yue, Franck Gallardo, Luisa Bustamante-Jaramillo, Gustaf E. Rydell, Maria Soller, Oliver Josef Müller, Marie-Lise Blondot, Qingxin Chen, Tomas Edreira
article en
2 citations

Abstract

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.

iScienceVol. 28(6)
Centre National de la Recherche Scientifique (FR), Université de Bordeaux (FR), German Cancer Research Center (DE), Heidelberg University (DE), Laboratoire de Microbiologie Fondamentale et Pathogénicité (FR), University of Gothenburg (SE)
Göteborgs Universitet
Openalex Percentile: Top 8%
Virus-based gene therapy research
3.63
DOIOpen Access PDFOpenAlex
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