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Targeted rearrangement of a chromosomal repeat sequence by transfection of a homologous DNA sequence using purified integrase

Gene Therapy volume 12, pages 783–794 (2005)Cite this article

Abstract

Using a liposomal transfection with purified bovine leukemia virus (BLV) integrase, we observed an efficient DNA rearrangement of a chromosomal repeat sequence and targeted integration of a part of the transfected plasmid. The BLV integrase recognition sequence (IRS) including the 3′ end of the BLV LTR U5, one of the sites cleaved by the integrase, was essential for the DNA rearrangement, and a sequence homologous to the chromosomal DNA neighboring the repeat target site had to be placed downstream of the IRS on the transfected plasmid. The pSV2neo DNA, including the pBR322 sequence preintegrated into L929 cells (primary transfectants), was rearranged by a secondary transfection of a pBR322-based hygromycin-resistance plasmid carrying the IRS. We present a model to explain the chromosomal DNA rearrangement of the primary clones through a homologous recombination-like reaction and amplification of the neighboring sequences.

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Acknowledgements

We thank Professor H Koyama of the Kihara Institute for Biological Research at Yokohama City University for critical reading and helpful discussions. We also thank Mr C Moreby for preparation of the manuscript. This work was supported in part by a grant from the Foundation of Human Health Sciences.

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  1. Department of Safety Research on Biologics and Blood Products, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo, Japan

    A S Tanaka & K Komuro

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  1. A S Tanaka
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Tanaka, A., Komuro, K. Targeted rearrangement of a chromosomal repeat sequence by transfection of a homologous DNA sequence using purified integrase. Gene Ther 12, 783–794 (2005). https://doi.org/10.1038/sj.gt.3302458

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