Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors
Abstract Recombinant adeno-associated viral vectors (rAAV) are a powerful tool for gene delivery but have a limited DNA carrying capacity. Efforts to expand this genetic payload have focused on engineering the vector components, such as dual trans-splicing vectors which double the delivery size by e...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-02-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-56738-5 |
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| _version_ | 1850197863087210496 |
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| author | Anna C. Maurer Brian Benyamini Oscar N. Whitney Vinson B. Fan Claudia Cattoglio Djem U. Kissiov Gina M. Dailey Xavier Darzacq Matthew D. Weitzman Robert Tjian |
| author_facet | Anna C. Maurer Brian Benyamini Oscar N. Whitney Vinson B. Fan Claudia Cattoglio Djem U. Kissiov Gina M. Dailey Xavier Darzacq Matthew D. Weitzman Robert Tjian |
| author_sort | Anna C. Maurer |
| collection | DOAJ |
| description | Abstract Recombinant adeno-associated viral vectors (rAAV) are a powerful tool for gene delivery but have a limited DNA carrying capacity. Efforts to expand this genetic payload have focused on engineering the vector components, such as dual trans-splicing vectors which double the delivery size by exploiting the natural concatenation of rAAV genomes in host nuclei. We hypothesized that inefficient dual vector transduction could be improved by modulating host factors which affect concatenation. Since factors mediating concatenation are not well defined, we performed a genome-wide screen to identify host cell regulators. We discover that Homologous Recombination (HR) is inhibitory to dual vector transduction. We demonstrate that depletion or inhibition of HR factors BRCA1 and Rad51 significantly increase reconstitution of a large split transgene by increasing both concatenation and expression from rAAVs. Our results define roles for DNA damage repair in rAAV transduction and highlight the potential for pharmacological intervention to increase genetic payload of rAAV vectors. |
| format | Article |
| id | doaj-art-efbba43e125e40ec8bf6ba10728fdc5c |
| institution | OA Journals |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-efbba43e125e40ec8bf6ba10728fdc5c2025-08-20T02:13:02ZengNature PortfolioNature Communications2041-17232025-02-0116111410.1038/s41467-025-56738-5Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectorsAnna C. Maurer0Brian Benyamini1Oscar N. Whitney2Vinson B. Fan3Claudia Cattoglio4Djem U. Kissiov5Gina M. Dailey6Xavier Darzacq7Matthew D. Weitzman8Robert Tjian9Department of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Molecular and Cell Biology, University of CaliforniaDepartment of Pathology and Laboratory Medicine, University of Pennsylvania Perelman School of Medicine and the Children’s Hospital of PhiladelphiaDepartment of Molecular and Cell Biology, University of CaliforniaAbstract Recombinant adeno-associated viral vectors (rAAV) are a powerful tool for gene delivery but have a limited DNA carrying capacity. Efforts to expand this genetic payload have focused on engineering the vector components, such as dual trans-splicing vectors which double the delivery size by exploiting the natural concatenation of rAAV genomes in host nuclei. We hypothesized that inefficient dual vector transduction could be improved by modulating host factors which affect concatenation. Since factors mediating concatenation are not well defined, we performed a genome-wide screen to identify host cell regulators. We discover that Homologous Recombination (HR) is inhibitory to dual vector transduction. We demonstrate that depletion or inhibition of HR factors BRCA1 and Rad51 significantly increase reconstitution of a large split transgene by increasing both concatenation and expression from rAAVs. Our results define roles for DNA damage repair in rAAV transduction and highlight the potential for pharmacological intervention to increase genetic payload of rAAV vectors.https://doi.org/10.1038/s41467-025-56738-5 |
| spellingShingle | Anna C. Maurer Brian Benyamini Oscar N. Whitney Vinson B. Fan Claudia Cattoglio Djem U. Kissiov Gina M. Dailey Xavier Darzacq Matthew D. Weitzman Robert Tjian Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors Nature Communications |
| title | Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors |
| title_full | Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors |
| title_fullStr | Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors |
| title_full_unstemmed | Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors |
| title_short | Double-strand break repair pathways differentially affect processing and transduction by dual AAV vectors |
| title_sort | double strand break repair pathways differentially affect processing and transduction by dual aav vectors |
| url | https://doi.org/10.1038/s41467-025-56738-5 |
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