Generating knockout rats by transposon mutagenesis in spermatogonial stem cells

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Abstract

Disrupting genes in the rat on a genome-wide scale will allow the investigation of many biological processes linked to human health. Here we used transposon-mediated mutagenesis to knock out genes in rat spermatogonial stem cells. Given the capacity of the testis to support spermatogenesis from thousands of transplanted, genetically manipulated spermatogonia, this approach paves a way for high-throughput functional genomic studies in the laboratory rat.

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Figure 1: Production of knockout rats using polyclonal libraries of mutant spermatogonia.
Figure 2: Production of knockout rats using monoclonally enriched spermatogonial lines.

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Acknowledgements

We thank T. Nguyen, T.E. Richardson, G. Mendrano and L.M. Thompson for help with these studies, N. Hübner, D.J. Mangelsdorf and M.H. Cobb for discussions and for critical reading of the manuscript, and EURATools–EURATrans consortium for sponsoring Z. Izsvák and F.K.H to attend annual meetings (grant HEALTH-F4-2010-241504). This work was supported by US National Institutes of Health grants R21RR023958 from the National Center for Research Resources and RO1HD036022 from the National Institute of Child Health and Human Development to F.K.H., by the Bundesministerium fur Bildung und Forschung (grant NGFN-2) to Z. Ivics, European Young Investigator award to Z. Izsvák and by the Cecil H. and Ida Green Center for Reproductive Biology Sciences at University of Texas Southwestern Medical Center in Dallas.

Author information

I.G., J.F., H.M.P., J.R.S., K.M.C. and F.K.H. performed the research; Z. Izsvák, Z. Ivics and F.K.H. designed research; all authors analyzed the data; Z. Izsvák, Z. Ivics and F.K.H. wrote the manuscript. Correspondence should be addressed to F.K.H. regarding spermatogonial technology or Z. Ivics regarding transposon technology.

Correspondence to Zoltán Ivics or F Kent Hamra.

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The authors declare no competing financial interests.

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Supplementary Figures 1–8, Supplementary Tables 1–5 and Supplementary Discussion (PDF 2237 kb)

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Izsvák, Z., Fröhlich, J., Grabundzija, I. et al. Generating knockout rats by transposon mutagenesis in spermatogonial stem cells. Nat Methods 7, 443–445 (2010) doi:10.1038/nmeth.1461

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