Small scale production of recombinant adeno-associated viral vectors for gene delivery to the nervous system

Joost Verhaagen, Barbara Hobo, Erich M.E. Ehlert, Ruben Eggers, Joanna A. Korecka, Stefan A. Hoyng, Callan L. Attwell, Alan R. Harvey, Matthew R.J. Mason

Research output: Chapter in Book/Conference paperChapter

2 Citations (Scopus)

Abstract

Adeno-associated viral vectors have numerous applications in neuroscience, including the study of gene function in health and disease, targeting of light-sensitive proteins to anatomically distinct sets of neurons to manipulate neuronal activity (optogenetics), and the delivery of fluorescent protein to study anatomical connectivity in the brain. Moreover several phase I/II clinical trials for gene therapy of eye and brain diseases with adeno-associated viral vectors have shown that these vectors are well tolerated by human patients. In this chapter we describe a detailed protocol for the small scale production of recombinant adeno-associated viral vectors. This protocol can be executed by investigators with experience in cell culture and molecular biological techniques in any well-equipped molecular neurobiology laboratory. With this protocol we typically obtain research batches of 100–200 μL that range in titer from 5 × 1012 to 2 × 1013 genomic copies/mL.

Original languageEnglish
Title of host publicationRetinal Gene Therapy
Subtitle of host publicationMethods and Protocols
EditorsC Boon, J Wijnholds
PublisherHumana Press
Pages3-17
Number of pages15
Volume1715
ISBN (Electronic)9781493975228
ISBN (Print)9781493975211
DOIs
Publication statusPublished - 1 Jan 2018

Publication series

NameMethods in Molecular Biology
Volume1715
ISSN (Print)1064-3745

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Verhaagen, J., Hobo, B., Ehlert, E. M. E., Eggers, R., Korecka, J. A., Hoyng, S. A., ... Mason, M. R. J. (2018). Small scale production of recombinant adeno-associated viral vectors for gene delivery to the nervous system. In C. Boon, & J. Wijnholds (Eds.), Retinal Gene Therapy: Methods and Protocols (Vol. 1715, pp. 3-17). (Methods in Molecular Biology; Vol. 1715). Humana Press. https://doi.org/10.1007/978-1-4939-7522-8_1