Retroviruses, acting as somatic cell insertional mutagens, have been widely used to identify cancer genes in the haematopoietic system and mammary gland1,2. An insertional mutagen for use in other mouse somatic cells would facilitate the identification of genes involved in tumour formation in a wider variety of tissues. Here we report the ability of the Sleeping Beauty transposon to act as a somatic insertional mutagen to identify genes involved in solid tumour formation. A Sleeping Beauty transposon, engineered to elicit loss-of-function or gain-of-function mutations, transposed in all somatic tissues tested and accelerated tumour formation in mice predisposed to cancer. Cloning transposon insertion sites from these tumours revealed the presence of common integration sites, at known and candidate cancer genes, similar to those observed in retroviral mutagenesis screens. Sleeping Beauty is a new tool for unbiased, forward genetic screens for cancer genes in vivo.
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We thank M. Roussel for providing Arf-/- mice; P. Woll, S. Rosenthal, R. Wang, P. Lobitz, J. Zinggeler and M. Davies for technical assistance; P. Hackett, S. McIvor, S. Ekker, P. Marker and members of the Largaespada laboratory for critical reading of the manuscript; N. Kirchhof for performing the pathology; T. L. Bergemann for providing assistance with statistical analysis; and K. Akagi for performing automated sequence analysis and BLAT searches. T2/Onc transgenic mice were generated by the University of Minnesota Mouse Genetics Laboratory. This work was supported by the Arnold and Mabel Beckman Foundation, the National Institute of Drug Abuse and the National Cancer Institute.
D.A.L. is a founder of Discovery Genomics, Inc. (DGI), a biotechnology company that has licensed SB technology from the University of Minnesota for use in human gene therapy. None of the work reported in this manuscript involved DGI.
Primer sequences. (DOC 22 kb)
Details of integrations in CISs. (DOC 366 kb)
Several T2/onc integrations occurred near previously identified leukaemia retroviral CISs. (DOC 20 kb)
T2/onc Southern analysis. (PDF 143 kb)
T2/onc integrations are tumour specific. (PDF 68 kb)
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