Adeno-Associated Virus Vector-Mediated Transgene Integration into Neurons and Other Nondividing Cell Targets

Author:

Wu Ping1,Phillips M. Ian2,Bui John2,Terwilliger Ernest F.1

Affiliation:

1. Divisions of Experimental Medicine and Hematology/Oncology, Beth Israel Deaconess Medical Center and Harvard Institutes of Medicine, Boston, Massachusetts 02115,1 and

2. Department of Physiology, College of Medicine, University of Florida, Gainesville, Florida 326102

Abstract

ABSTRACT The site-specific integration of wild-type adeno-associated virus (wtAAV) into the human genome is a very attractive feature for the development of AAV-based gene therapy vectors. However, knowledge about integration of wtAAV, as well as currently configured recombinant AAV (rAAV) vectors, is limited. By using a modified Alu-PCR technique to amplify and sequence the vector-cellular junctions, we provide the first direct evidence both in vitro and in vivo of rAAV-mediated transgene integration in several types of nondividing cells, including neurons. This novel technique will be highly useful for further delineating the mechanisms underlying AAV-mediated integration, including issues of frequency, site preference, and DNA rearrangement in human as well as animal cells. Results from these studies should be beneficial for the development of the next generation of gene delivery vectors.

Publisher

American Society for Microbiology

Subject

Virology,Insect Science,Immunology,Microbiology

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