Mammalian mutagenesis using a highly mobile somatic Sleeping Beauty transposon system

Abstract

Transposons have provided important genetic tools for functional genomic screens in lower eukaryotes but have proven less useful in higher eukaryotes because of their low transposition frequency. Here we show that Sleeping Beauty (SB), a member of the Tc1/mariner class of transposons, can be mobilized in mouse somatic cells at frequencies high enough to induce embryonic death and cancer in wild-type mice. Tumours are aggressive, with some animals developing two or even three different types of cancer within a few months of birth. The tumours result from SB insertional mutagenesis of cancer genes, thus facilitating the identification of genes and pathways that induce disease. SB transposition can easily be controlled to mutagenize any target tissue and can therefore, in principle, be used to induce many of the cancers affecting humans, including those for which little is known about the aetiology. The uses of SB are also not restricted to the mouse and could potentially be used for forward genetic screens in any higher eukaryote in which transgenesis is possible.

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Figure 1: Analysis of double-transgenic embryos and adults.
Figure 2: Adult double-transgenic mice die from cancer.
Figure 3: Medulloblastoma pathology.
Figure 4: Analysis of Notch1 integrations.
Figure 5: Notch1 cooperating genes.

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Acknowledgements

We thank D. Swing and R. Koogle for generating the T2/Onc2 transgenic mice and maintaining all mouse strains, and E. Southon and S. Reed for generating mice carrying the RosaSB knock-in allele. This research is supported by the Department of Health and Human Services, National Institutes of Health and the National Cancer Institute.

Author information

Correspondence to Nancy A. Jenkins.

Ethics declarations

Competing interests

Dr Largaespada is a cofounder of Discovery Genomics Inc., which has licensed Sleeping Beauty technology from the University of Minnesota for applications unrelated to this work.

Supplementary information

Supplementary Figures S1 and S2

Supplementary Figure S1 shows generation and characterization of T2/Onc2 transgenic founders. Supplementary Figure S2 shows generation and characterization of RosaSB knock-in allele (PPT 2426 kb)

Supplementary Methods

Describes Southerns, Northerns and LM-PCR protocol (PDF 97 kb)

Supplementary Table S1

Lists all annotation data collected by cloning transposon junctions from double transgenic embryos (XLS 85 kb)

Supplementary Table S2

Lists all common sites of transposon integration identified in the data described in Supplementary Table S1 (XLS 18 kb)

Supplementary Table S3

Lists all annotation data collected by cloning transposon junctions from tumors (XLS 138 kb)

Supplementary Table S4

Lists all common sites of transposon integration identified in the data described in Supplementary Table S3 (XLS 33 kb)

Supplementary Table S5

Demonstrates the transposon orientation bias seen in the tumor data when compared with the embryo data (XLS 19 kb)

Supplementary Table S6

Lists pathways commonly affected by transposon integration in tumors (XLS 17 kb)

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