Small Scale Production of Recombinant Adeno-Associated Viral Vectors for Gene Delivery to the Nervous System

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

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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
Pages (from-to)3-17
Number of pages15
JournalMethods in Molecular Biology
Volume1715
DOIs
Publication statusPublished - 03 Jan 2018

Keywords

  • Journal Article

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