| Chapter title |
Production and Characterization of Vectors Based on the Cardiotropic AAV Serotype 9.
|
|---|---|
| Chapter number | 6 |
| Book title |
Cardiac Gene Therapy
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6588-5_6 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6586-1, 978-1-4939-6588-5
|
| Authors |
Erik Kohlbrenner, Thomas Weber Ph.D., Thomas Weber |
| Editors |
Kiyotake Ishikawa |
| Abstract |
Vectors based on adeno-associated virus serotype 9 (AAV9) efficiently transduce cardiomyocytes in both rodents and large animal models upon either systemic or regional vector delivery. In this chapter, we describe the most widely used production and purification method of AAV9. This production approach does not depend on the use of a helpervirus but instead on transient transfection of HEK293T cells with a plasmid containing the recombinant AAV genome and a second plasmid encoding the AAV9 capsid proteins, the AAV Rep proteins and the adenoviral helper functions. The recombinant AAV is then purified by iodixanol density gradient centrifugation. This chapter also describes in detail the characterization and quality control methods required for assuring high quality vector preparations, which is of particular importance for experiments in large animal models. |
Mendeley readers
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 35 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 6 | 17% |
| Researcher | 6 | 17% |
| Student > Bachelor | 4 | 11% |
| Professor > Associate Professor | 2 | 6% |
| Student > Master | 2 | 6% |
| Other | 3 | 9% |
| Unknown | 12 | 34% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 6 | 17% |
| Pharmacology, Toxicology and Pharmaceutical Science | 4 | 11% |
| Chemical Engineering | 4 | 11% |
| Agricultural and Biological Sciences | 3 | 9% |
| Immunology and Microbiology | 2 | 6% |
| Other | 4 | 11% |
| Unknown | 12 | 34% |