Small and Micro-Scale Recombinant Adeno-Associated Virus Production and Purification for Ocular Gene Therapy Applications. Methods Mol Biol 2018;1715:19-31
Date
12/01/2017Pubmed ID
29188503DOI
10.1007/978-1-4939-7522-8_2Scopus ID
2-s2.0-85036634077 (requires institutional sign-in at Scopus site) 15 CitationsAbstract
Over the past two decades recombinant adeno-associated virus (rAAV) vectors have emerged as the gold standard for transferring genetic material to cells of the retina. The ability to effectively produce small batches of rAAV vector at high enough purity for in vitro and in vivo applications in a cost-effective manner is paramount. This is particularly the case when conducting preclinical experiments to screen novel serotypes, promoters or transgenes, where production of numerous vector batches is required. Current vector production methods often produce large quantities of vector, limiting the cost-effectiveness and practicality of such screening experiments, which often require only small volumes of vector to carry out. Herein, we describe a method to produce high titer (1012-1013 vector genomes (vg)/mL) rAAV vector on small (~100 μL) or micro (~15 μL) scale for in vitro and in vivo applications.
Author List
Reid CA, Lipinski DMAuthor
Daniel M. Lipinski PhD Associate Professor in the Ophthalmology and Visual Sciences department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
DependovirusGene Transfer Techniques
Genetic Therapy
Genetic Vectors
HEK293 Cells
Humans
Plasmids
Retina
