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

TY - JOUR

T1 - Small Scale Production of Recombinant Adeno-Associated Viral Vectors for Gene Delivery to the Nervous System

AU - Verhaagen, J.

AU - Hobo, B.

AU - Ehlert, Erich M E

AU - Eggers, R.

AU - Korecka, Joanna A

AU - Hoyng, Stefan A

AU - Attwell, Callan L

AU - Harvey, Alan R

AU - Mason, Matthew R J

PY - 2018/1/3

Y1 - 2018/1/3

N2 - 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.

AB - 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.

KW - Journal Article

U2 - 10.1007/978-1-4939-7522-8_1

DO - 10.1007/978-1-4939-7522-8_1

M3 - Article

C2 - 29188502

SN - 1064-3745

VL - 1715

SP - 3

EP - 17

JO - Methods in Molecular Biology

JF - Methods in Molecular Biology

ER -