High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector

The successful application of recombinant adeno-associated virus (rAAV) vectors for long-term transgene expression in clinical studies requires scalable production methods with genetically stable components. Due to their simple production scheme and the high viral titers achievable, first generation...

Full description

Bibliographic Details
Main Author: Stefan Weger
Format: Article
Language:English
Published: MDPI AG 2022-12-01
Series:Viruses
Subjects:
Online Access:https://www.mdpi.com/1999-4915/15/1/64
_version_ 1797436515412869120
author Stefan Weger
author_facet Stefan Weger
author_sort Stefan Weger
collection DOAJ
description The successful application of recombinant adeno-associated virus (rAAV) vectors for long-term transgene expression in clinical studies requires scalable production methods with genetically stable components. Due to their simple production scheme and the high viral titers achievable, first generation recombinant adenoviruses (rAdV) have long been taken into consideration as suitable tools for simultaneously providing both the helper functions and the AAV <i>rep</i> and <i>cap</i> genes for rAAV packaging. So far, however, such rAdV-<i>rep/cap</i> vectors have been difficult to generate and often turned out to be genetically unstable. Through ablation of <i>cis</i> and <i>trans</i> inhibitory function in the AAV-2 genome we have succeeded in establishing separate and stable rAdVs for high-level AAV serotype 2 Rep and Cap expression. These allowed rAAV-2 production at high burst sizes by simple coinfection protocols after providing the AAV-ITR flanked transgene vector genome either as rAAV-2 particles at low input concentrations or in form of an additional rAdV. With characteristics such as the ease of producing the required components, the straightforward adaption to other transgenes and the possible extension to further serotypes or capsid variants, especially the rAdV-mediated rAAV amplification system presents a very promising candidate for up-scaling to clinical grade vector preparations.
first_indexed 2024-03-09T11:03:44Z
format Article
id doaj.art-f9328b85c77843f483ba0ad162cd8a6a
institution Directory Open Access Journal
issn 1999-4915
language English
last_indexed 2024-03-09T11:03:44Z
publishDate 2022-12-01
publisher MDPI AG
record_format Article
series Viruses
spelling doaj.art-f9328b85c77843f483ba0ad162cd8a6a2023-12-01T01:07:43ZengMDPI AGViruses1999-49152022-12-011516410.3390/v15010064High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input VectorStefan Weger0Charité–Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Clinic for Neurology with Experimental Neurology, Gene Therapy Group, Campus Benjamin Franklin, Hindenburgdamm27, 12203 Berlin, GermanyThe successful application of recombinant adeno-associated virus (rAAV) vectors for long-term transgene expression in clinical studies requires scalable production methods with genetically stable components. Due to their simple production scheme and the high viral titers achievable, first generation recombinant adenoviruses (rAdV) have long been taken into consideration as suitable tools for simultaneously providing both the helper functions and the AAV <i>rep</i> and <i>cap</i> genes for rAAV packaging. So far, however, such rAdV-<i>rep/cap</i> vectors have been difficult to generate and often turned out to be genetically unstable. Through ablation of <i>cis</i> and <i>trans</i> inhibitory function in the AAV-2 genome we have succeeded in establishing separate and stable rAdVs for high-level AAV serotype 2 Rep and Cap expression. These allowed rAAV-2 production at high burst sizes by simple coinfection protocols after providing the AAV-ITR flanked transgene vector genome either as rAAV-2 particles at low input concentrations or in form of an additional rAdV. With characteristics such as the ease of producing the required components, the straightforward adaption to other transgenes and the possible extension to further serotypes or capsid variants, especially the rAdV-mediated rAAV amplification system presents a very promising candidate for up-scaling to clinical grade vector preparations.https://www.mdpi.com/1999-4915/15/1/64rAAV packagingadeno-associated virusrecombinant adenovirusRIS-Ad
spellingShingle Stefan Weger
High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector
Viruses
rAAV packaging
adeno-associated virus
recombinant adenovirus
RIS-Ad
title High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector
title_full High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector
title_fullStr High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector
title_full_unstemmed High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector
title_short High-Level rAAV Vector Production by rAdV-Mediated Amplification of Small Amounts of Input Vector
title_sort high level raav vector production by radv mediated amplification of small amounts of input vector
topic rAAV packaging
adeno-associated virus
recombinant adenovirus
RIS-Ad
url https://www.mdpi.com/1999-4915/15/1/64
work_keys_str_mv AT stefanweger highlevelraavvectorproductionbyradvmediatedamplificationofsmallamountsofinputvector