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Nucleic Acids Research Advance Access originally published online on October 25, 2008
Nucleic Acids Research 2008 36(22):e148; doi:10.1093/nar/gkn801
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Nucleic Acids Research, 2008, Vol. 36, No. 22 e148
© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Bacteriophage Mu integration in yeast and mammalian genomes

Anja O. Paatero1, Hilkka Turakainen1, Lotta J. Happonen1, Cia Olsson2, Tiina Palomäki2, Maria I. Pajunen1,3, Xiaojuan Meng4,{dagger}, Timo Otonkoski2,5, Timo Tuuri2,6, Charles Berry7, Nirav Malani7, Mikko J. Frilander4, Frederic D. Bushman7 and Harri Savilahti1,3,*

1Program in Cellular Biotechnology, Institute of Biotechnology, Viikki Biocenter, 2Biomedicum Stem Cell Center, Biomedicum Helsinki, University of Helsinki, Helsinki, 3Division of Genetics and Physiology, Department of Biology, University of Turku, Turku, 4Program in Developmental Biology, Institute of Biotechnology, Viikki Biocenter, University of Helsinki, 5Hospital for Children and Adolescents, University of Helsinki, 6Family Federation of Finland, Helsinki, Finland and 7Department of Microbiology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA

*To whom correspondence should be addressed. Tel: +358 2 333 5586; Fax: +358 2 333 6680; Email: harri.savilahti{at}utu.fi

Received August 26, 2008. Revised October 9, 2008. Accepted October 10, 2008.

Genomic parasites have evolved distinctive lifestyles to optimize replication in the context of the genomes they inhabit. Here, we introduced new DNA into eukaryotic cells using bacteriophage Mu DNA transposition complexes, termed ‘transpososomes’. Following electroporation of transpososomes and selection for marker gene expression, efficient integration was verified in yeast, mouse and human genomes. Although Mu has evolved in prokaryotes, strong biases were seen in the target site distributions in eukaryotic genomes, and these biases differed between yeast and mammals. In Saccharomyces cerevisiae transposons accumulated outside of genes, consistent with selection against gene disruption. In mouse and human cells, transposons accumulated within genes, which previous work suggests is a favorable location for efficient expression of selectable markers. Naturally occurring transposons and viruses in yeast and mammals show related, but more extreme, targeting biases, suggesting that they are responding to the same pressures. These data help clarify the constraints exerted by genome structure on genomic parasites, and illustrate the wide utility of the Mu transpososome technology for gene transfer in eukaryotic cells.

Present address: Charles Berry, Department of Family and Preventive Medicine, School of Medicine, University of California San Diego, La Jolla, CA, USA

{dagger}Deceased

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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