Adeno-associated virus Type 2 Rep proteins mediate integration of lentiviral vectors
- Published
- Accepted
- Subject Areas
- Biotechnology, Molecular Biology, Virology
- Keywords
- Integration, Adeno-associated virus, Lentivirus Vectors, Rep proteins
- Licence
- This is an open access article, free of all copyright, made available under the Creative Commons Public Domain Dedication. This work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
- Cite this article
- 2014. Adeno-associated virus Type 2 Rep proteins mediate integration of lentiviral vectors. PeerJ PrePrints 2:e326v4 https://doi.org/10.7287/peerj.preprints.326v4
Abstract
Aims: Adeno-associated virus type 2 (AAV2) is a naturally defective human parvovirus that is being developed as a gene therapy vector. In dividing cells, AAV2 DNA persists by integration into the host chromosomes. AAV2 is unique among mammalian viruses in its ability to integrate preferentially into a particular locus within human chromosome 19, designated AAVS1(also known as Mbs 85). The AAV2 Rep68 and Rep78 proteins mediate this integration. Recent data suggest that Rep68 and Rep78 can mediate integration of non-AAV2 DNA with free ends. To test this hypothesis, we targeted insertion of different lentiviral vectors to AAVS1. Methods: Cells were co-infected with wild-type AAV2, and integrase-proficient or integrase-deficient lentivirus vectors. A highly specific PCR-based assay was used to detect lentivirus integration at AAVS1. Similar experiments were performed using lentiviral vectors containing the AAV2 rep gene. Results: All lentiviral vectors tested integrated at AAVS1, if the rep gene was present either within the lentiviral vector or supplied in trans. All that was required for integration at AAVS1 was the amino acid sequence shared between Rep68 and Rep78. The results were similar with integrase-proficient or integrase-deficient lentiviral vectors. Conclusions: The inclusion of the rep gene with lentiviral vectors may produce more predictable integration patterns.
Author Comment
This version contains a typographical error correction.