1. Generation of infectious recombinant Adeno-associated virus in Saccharomyces cerevisiae.
- Author
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Barajas, Daniel, Aponte-Ubillus, Juan Jose, Akeefe, Hassibullah, Cinek, Tomas, Peltier, Joseph, and Gold, Daniel
- Subjects
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ADENO-associated virus , *RECOMBINANT viruses , *SACCHAROMYCES cerevisiae , *VIRUS diseases , *GENE delivery techniques , *EUKARYOTES - Abstract
The yeast Saccharomyces cerevisiae has been successfully employed to establish model systems for a number of viruses. Such model systems are powerful tools to study the virus biology and in particular for the identification and characterization of host factors playing a role in the viral infection cycle. Adeno-associated viruses (AAV) are heavily studied due to their use as gene delivery vectors. AAV relies on other helper viruses for successful replication and on host factors for several aspects of the viral life cycle. However the role of host and helper viral factors is only partially known. Production of recombinant AAV (rAAV) vectors for gene delivery applications depends on knowledge of AAV biology and the limited understanding of host and helper viral factors may be precluding efficient production, particularly in heterologous systems. Model systems in simpler eukaryotes like the yeast S. cerevisiae would be useful tools to identify and study the role of host factors in AAV biology. Here we show that expression of AAV2 viral proteins VP1, VP2, VP3, AAP, Rep78, Rep52 and an ITR-flanked DNA in yeast leads to capsid formation, DNA replication and encapsidation, resulting in formation of infectious particles. Many of the AAV characteristics observed in yeast resemble those in other systems, making it a suitable model system. Future findings in the yeast system could be translatable to other AAV host systems and aid in more efficient production of rAAV vectors. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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