| Chapter title |
Small Scale Production of Recombinant Adeno-Associated Viral Vectors for Gene Delivery to the Nervous System
|
|---|---|
| Chapter number | 1 |
| Book title |
Retinal Gene Therapy
|
| Published in |
Methods in molecular biology, January 2018
|
| DOI | 10.1007/978-1-4939-7522-8_1 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-7521-1, 978-1-4939-7522-8
|
| Authors |
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 |
| 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. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 31 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 10 | 32% |
| Student > Master | 5 | 16% |
| Student > Ph. D. Student | 4 | 13% |
| Student > Bachelor | 2 | 6% |
| Professor | 1 | 3% |
| Other | 1 | 3% |
| Unknown | 8 | 26% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 6 | 19% |
| Agricultural and Biological Sciences | 5 | 16% |
| Neuroscience | 5 | 16% |
| Psychology | 1 | 3% |
| Medicine and Dentistry | 1 | 3% |
| Other | 1 | 3% |
| Unknown | 12 | 39% |