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Optimization of viral protein ratios for production of rAAV serotype 5 in the baculovirus system
- Source :
- Gene Therapy. 25:415-424
- Publication Year :
- 2018
- Publisher :
- Springer Science and Business Media LLC, 2018.
-
Abstract
- Recombinant adeno-associated virus (rAAV) has become the vector of choice for the development of novel human gene therapies. High-yield manufacturing of high-quality vectors can be achieved using the baculovirus expression vector system. However, efficient production of rAAV in this insect cell-based system requires a genetic redesign of the viral protein 1 (VP1) operon. In this study, we generated a library of rationally designed rAAV serotype 5 variants with modulations in the translation-initiation region of VP1 and investigated the potency of the resulting vectors. We found that the initiation strength at the VP1 translational start had downstream effects on the VP2/VP3 ratio. Excessive incorporation of VP3 into a vector type decreased potency, even when the VP1/VP2 ratio was in balance. Finally, we successfully generated a potent rAAV vector based on serotype 5 with a balanced VP1/VP2/VP3 stoichiometry.
- Subjects :
- 0301 basic medicine
Serotype
Baculoviridae
Operon
Viral protein
viruses
Genetic Vectors
Serogroup
medicine.disease_cause
Virus
law.invention
Viral Proteins
03 medical and health sciences
0302 clinical medicine
Parvovirinae
law
Genetics
medicine
Humans
Vector (molecular biology)
Molecular Biology
Gene
biology
virus diseases
Genetic Therapy
Dependovirus
biochemical phenomena, metabolism, and nutrition
biology.organism_classification
Virology
030104 developmental biology
030220 oncology & carcinogenesis
Recombinant DNA
Molecular Medicine
Capsid Proteins
Subjects
Details
- ISSN :
- 14765462 and 09697128
- Volume :
- 25
- Database :
- OpenAIRE
- Journal :
- Gene Therapy
- Accession number :
- edsair.doi.dedup.....7064631ccf19d58b808f9b41259c53e2