S/MAR Element Facilitates Episomal Long-Term Persistence of Adeno-Associated Virus Vector Genomes in Proliferating Cells
| Autor: | Philip Boehme, Hans J. Lipps, Maria Schnödt-Fuchs, Claudia Hagedorn, Heba Abdelrazik, Hildegard Büning |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
viruses Transgene Genetic enhancement Population Genetic Vectors Genome Viral Biology medicine.disease_cause Genome Virus 03 medical and health sciences 0302 clinical medicine Genetics medicine Humans Vector (molecular biology) education Scaffold/matrix attachment region Molecular Biology Adeno-associated virus education.field_of_study Dependovirus Matrix Attachment Regions Virology 030104 developmental biology HEK293 Cells 030220 oncology & carcinogenesis Molecular Medicine HeLa Cells Plasmids |
| Zdroj: | Human gene therapy. 28(12) |
| ISSN: | 1557-7422 |
| Popis: | Adeno-associated virus (AAV) vectors are one of the most frequently applied gene transfer systems in research and human clinical trials. Since AAV vectors do not possess an integrase activity, application is restricted to terminally differentiated tissues if transgene expression is required long term. To overcome this limitation and to generate AAV vectors that persist episomally in dividing cells, AAV vector genomes were equipped with a scaffold/matrix attachment region (S/MAR). After a mild antibiotic selection, cells transduced with AAV-S/MAR established colonies that maintained long-term transgene expression (50 population doublings) from replicating AAV vector episomes in the absence of further selection. Unexpectedly, with a lesser but still significant efficiency, the control vector (AAV-ΔS/MAR), a standard single-stranded AAV vector, also established stable transgene-expressing colonies, most of which were maintained as replicating episomes rather than integrated vector genomes. Thus, based on the result in HeLa cells, it is concluded that AAV vector genomes per se possess the ability to establish episomal maintenance in proliferating cells, a feature that can be enhanced by incorporation of a foreign genomic element such as an S/MAR element. |
| Databáze: | OpenAIRE |
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