Cancer gene discovery in solid tumours using transposon-based somatic mutagenesis in the mouse

Abstract

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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Figure 1: Vector design and somatic transposition.
Figure 2: Arf -/- ;T2/Onc;CAGGS-SB10 mice have a shorter tumour latency than singly transgenic controls.
Figure 3: Activation of Braf by T2/Onc insertion.

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Acknowledgements

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.

Author information

Correspondence to David A. Largaespada.

Ethics declarations

Competing interests

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.

Supplementary information

Supplementary Methods

Primer sequences. (DOC 22 kb)

Supplementary Table S1

Details of integrations in CISs. (DOC 366 kb)

Supplementary Table S2

Several T2/onc integrations occurred near previously identified leukaemia retroviral CISs. (DOC 20 kb)

Supplementary Figure S1

T2/onc Southern analysis. (PDF 143 kb)

Supplementary Figure S2

T2/onc integrations are tumour specific. (PDF 68 kb)

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