Strategies to optimize capsid protein expression and single-stranded DNA formation of adeno-associated virus in<i>Saccharomyces cerevisiae</i>

Galli, Alvaro; Latta, Veronica Della; Bologna, Cinzia; Pucciarelli, Daniela; Cipriani, Filippo; Backovic, Ana; Cervelli, Tiziana

doi:10.1111/jam.13511

Cited by 9 publications

(12 citation statements)

References 61 publications

(81 reference statements)

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“… 6 , 7 The advantage of this system is that rep - cap sequence and rAAV genome are sequestered in different sub-cellular compartments, which eliminates the contamination with replication-competent AAV (rcAAV). Following seminal works of A. Galli’s 8 , 9 and R.O. Snyder’s groups, 10 , 11 Barajas et al.…”

Section: Main Textmentioning

confidence: 99%

Pharmacology of Recombinant Adeno-associated Virus Production

Penaud-Budloo

François

Clément

et al. 2018

Molecular Therapy - Methods & Clinical Development

139

131

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Recombinant adeno-associated viral (rAAV) vectors have been used in more than 150 clinical trials with a good safety profile and significant clinical benefit in many genetic diseases. In addition, due to their ability to infect non-dividing and dividing cells and to serve as efficient substrate for homologous recombination, rAAVs are being used as a tool for gene-editing approaches. However, manufacturing of these vectors at high quantities and fulfilling current good manufacturing practices (GMP) is still a challenge, and several technological platforms are competing for this niche. Herein, we will describe the most commonly used upstream methods to produce rAAVs, paying particular attention to the starting materials (input) used in each platform and which related impurities can be expected in final products (output). The most commonly found impurities in rAAV stocks include defective particles (i.e., AAV capsids that do contain the therapeutic gene or are not infectious), residual proteins from host cells and helper viruses (adenovirus, herpes simplex virus, or baculoviruses), and illegitimate DNA from plasmids, cells, or helper viruses that may be encapsidated into rAAV particles. Given the role that impurities may play in immunotoxicity, this article reviews the impurities inherently associated with each manufacturing platform.

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Section: Main Textmentioning

confidence: 99%

Pharmacology of Recombinant Adeno-associated Virus Production

Penaud-Budloo

François

Clément

et al. 2018

Molecular Therapy - Methods & Clinical Development

139

131

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“…mammalian and insect cells, 40 and possibly (although inefficiently) in yeast. 41,42 This implies that the host cellular factors required for AAV replication are conserved among eukaryotes.…”

Section: Contributions Of Cellular Protein Interactionsmentioning

confidence: 99%

Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction

Maurer

Weitzman

2020

Human Gene Therapy

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Recombinant adeno-associated virus has emerged as one of the most promising gene therapy delivery vectors. Development of these vectors took advantage of key features of the wild-type adeno-associated virus (AAV), enabled by basic studies of the underlying biology and requirements for transcription, replication, and packaging of the viral genome. Each step in generating and utilizing viral vectors involves numerous molecular interactions that together determine the efficiency of vector production and gene delivery. Once delivered into the cell, interactions with host proteins will determine the fate of the viral genome, and these will impact the intended goal of gene delivery. Here, we provide an overview of known interactions of the AAV genome with viral and cellular proteins involved in its amplification, packaging, and expression. Further appreciation of how the AAV genome interacts with host factors will enhance how this simple virus can be harnessed for an array of vector purposes that benefit human health.

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“…The potential development of a microbial platform would bring simple, lower‐cost, and scalable means for rAAV vector biomanufacturing. Proof‐of‐concept results showed efficient rAAV DNA replication and capsid formation, but low volumetric rAAV2 yield (Barajas et al, 2017; Galli et al, 2017). The low vector yields obtained in yeast could come as a result of an unbalanced expression of rAAV proteins, or a negative effect of host cell response to expression of foreign proteins and multiple forms of rAAV DNA.…”

Section: Introductionmentioning

confidence: 99%

Proteome profiling and vector yield optimization in a recombinant adeno‐associated virus‐producing yeast model

et al. 2020

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Strategies to optimize capsid protein expression and single-stranded DNA formation of adeno-associated virus inSaccharomyces cerevisiae

Abstract: New manufacturing strategies for AAV-based gene therapy vectors (rAAV) are needed to reduce costs and time of production. Our study explores the feasibility of yeast as alternative system for rAAV production.

Cited by 9 publications

References 61 publications

Pharmacology of Recombinant Adeno-associated Virus Production

Pharmacology of Recombinant Adeno-associated Virus Production

Adeno-Associated Virus Genome Interactions Important for Vector Production and Transduction

Proteome profiling and vector yield optimization in a recombinant adeno‐associated virus‐producing yeast model

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