MCB Accepts, published online ahead of print on 2 November 2009

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Mol. Cell. Biol. doi:10.1128/MCB.00913-09
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Tagged Mutagenesis by Efficient Minos based germ line transposition

Ton de Wit, Sylvia Dekker, Alex Maas, Guido Breedveld, Tobias A. Knoch, An Langeveld, Dorota Szumska, Roger Craig, Shoumo Bhattacharya, Frank Grosveld, and Dubravka Drabek^*

Department of Cell Biology, Erasmus Medical Center, Cancer Genomics Center (NGI, NL), POBOX 2040, 3000CA Rotterdam, The Netherlands; Cancer Genomics Center (NGI, NL), Department of Clinical Genetics, Erasmus Medical Center, POBOX 2040, 3000 CA Rotterdam, The Netherlands; The Wellcome Trust Centre for Human Genetics (WTCHG), the Henry Wellcome Building of Genomic Medicine, University of Oxford, UK; Minos Biosystems Ltd, Cheshire UK

^* To whom correspondence should be addressed. Email: d.drabek{at}erasmusmc.nl.

Germline gene transposition technology has been used to generate "libraries" of flies and worms carrying genome wide mutations. Phenotypic screening and DNA sequencing of such libraries provides functional information resulting from insertional events in target genes. There is also a great need to have a fast and efficient way to generate mouse mutants in vivo to model developmental defects and human diseases. Here we describe an optimized mammalian germ-line transposition system active during early mouse spermatogenesis using the Minos transposon. Transposon positive progeny carry on average more than 2 new transpositions and 45-100% of the progeny carry an insertion in a gene. The optimised Minos based system was tested in a small rapid dominant functional screen to identify mutated genes likely to cause measurable cardiovascular "disease" phenotypes in progeny/embryos. Importantly this system allows rapid screening for modifier genes.