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A mouse model of human L1 retrotransposition

Nature Genetics volume 32, pages 655–660 (2002)Cite this article

Abstract

The L1 retrotransposon has had an immense impact on the size and structure of the human genome through a variety of mechanisms, including insertional mutagenesis1,2. To study retrotransposition in a living organism, we created a mouse model of human L1 retrotransposition. Here we show that L1 elements can retrotranspose in male germ cells, and that expression of a human L1 element under the control of its endogenous promoter is restricted to testis and ovary. In the mouse line with the highest level of L1 expression, we found two de novo L1 insertions in 135 offspring. Both insertions were structurally indistinguishable from natural endogenous insertions. This suggests that an individual L1 element can have substantial mutagenic potential. In addition to providing a valuable in vivo model of retrotransposition in mammals, these mice are an important step in the development of a new random mutagenesis system.

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Figure 1: Transgenic mouse lines.
Figure 2: L1 transgene expression in germ cell fractions.
Figure 3: Expression from retrotransposition events in condensing-spermatid fractions.
Figure 4: PCR of germ-cell DNA to detect retrotransposition.
Figure 5: L1 insertions in transgenic mice.

References

  1. The International Human Genome Sequencing Consortium. Nature 409, 860–921 (2001).

  2. Ostertag, E.M. & Kazazian, H.H. Jr. Biology of mammalian L1 retrotransposons. Ann. Rev. Genet. 35, 501–538 (2001).

    CAS  Article  Google Scholar 

  3. Sassaman, D.M. et al. Many human L1 elements are capable of retrotransposition. Nature Genet. 16, 37–43 (1997).

    CAS  Article  Google Scholar 

  4. Moran, J.V. et al. High frequency retrotransposition in cultured mammalian cells. Cell 87, 917–927 (1996).

    CAS  Article  Google Scholar 

  5. Branciforte, D. & Martin, S.L. Developmental and cell type specificity of LINE-1 expression in mouse testis: implications for transposition. Mol. Cell. Biol. 14, 2584–2592 (1994).

    CAS  Article  Google Scholar 

  6. Trelogan, S.A. & Martin, S.L. Tightly regulated, developmentally specific expression of the first open reading frame from LINE-1 during mouse embryogenesis. Proc. Natl Acad. Sci. USA 92, 1520–1524 (1995).

    CAS  Article  Google Scholar 

  7. Nakanishi, T. et al. Real-time observation of acrosomal dispersal from mouse sperm using GFP as a marker protein. FEBS Lett. 449, 277–283 (1999).

    CAS  Article  Google Scholar 

  8. Ostertag, E.M. et al. Determination of L1 retrotransposition kinetics in cultured cells. Nucleic Acids Res. 28, 1418–1423 (2000).

    CAS  Article  Google Scholar 

  9. Schwahn, U. et al. Positional cloning of the gene for X-linked retinitis pigmentosa 2. Nature Genet. 19, 327–332 (1998).

    CAS  Article  Google Scholar 

  10. Kimberland, M.L. et al. Full-length human L1 insertions retain the capacity for high-frequency retrotransposition in cultured cells. Hum. Mol. Genet. 8, 1557–1560 (1999).

    CAS  Article  Google Scholar 

  11. Romrell, L.J., Bellvé, A.R. & Fawcett, D.W. Separation of mouse spermatogenic cells by sedimentation velocity. A morphological characterization. Dev. Biol. 49, 119–131 (1976).

    CAS  Article  Google Scholar 

  12. Welch, J.E., Schatte, E.C., O'Brien, D.A. & Eddy, E.M. Expression of a glyceraldehyde 3-phosphate dehydrogenase gene specific to mouse spermatogenic cells. Biol. Reprod. 46, 869–878 (1992).

    CAS  Article  Google Scholar 

  13. Steger, K. Transcriptional and translational regulation of gene expression in haploid spermatids. Anat. Embryol. 199, 471–487 (1999).

    CAS  Article  Google Scholar 

  14. Swergold, G.D. Identification, characterization, and cell specificity of a human LINE-1 promoter. Mol. Cell. Biol. 10, 6718–6729 (1990).

    CAS  Article  Google Scholar 

  15. Loeb, D.D. et al. The sequence of a large L1Md element reveals a tandemly repeated 5′ end and several features found in retrotransposons. Mol. Cell. Biol. 6, 168–182 (1986).

    CAS  Article  Google Scholar 

  16. Padgett, R.W., Hutchison, C.A. 3rd & Edgell, M.H. The F-type 5′ motif of mouse L1 elements: a major class of L1 termini similar to the A-type in organization but unrelated in sequence. Nucleic Acids Res. 16, 739–749 (1988).

    CAS  Article  Google Scholar 

  17. Naas, T.P. et al. An actively retrotranposing, novel subfamily of mouse L1 elements. EMBO J. 17, 590–597 (1998).

    CAS  Article  Google Scholar 

  18. Luan, D.D., Korman, M.H., Jakubczak, J.L. & Eickbush, T.H. Reverse transcription of R2Bm RNA is primed by a nick at the chromosomal target site: a mechanism for non-LTR retrotransposable elements. Cell 72, 595–605 (1993).

    CAS  Article  Google Scholar 

  19. Ostertag, E.M. & Kazazian, H.H. Jr. Twin Priming, a proposed mechanism for the creation of inversions in L1 retrotransposition. Genet. Res. 11, 2059–2065 (2001).

    CAS  Article  Google Scholar 

  20. Feng, Q., Moran, J.V., Kazazian, H.H. Jr. & Boeke, J.D. Human L1 retrotransposon encodes a conserved endonuclease required for retrotransposition. Cell 87, 905–916 (1996).

    CAS  Article  Google Scholar 

  21. Jurka, J. Sequence patterns indicate an enzymatic involvement in integration of mammalian retroposons. Proc. Natl Acad. Sci. USA 94, 1872–1877 (1997).

    CAS  Article  Google Scholar 

  22. Cost, G.J. & Boeke, J.D. Targeting of human retrotransposon integration is directed by the specificity of the L1 endonuclease for regions of unusual DNA structure. Biochem. 37, 18081–18093 (1998).

    CAS  Article  Google Scholar 

  23. Boissinot, S., Chevret, P. & Furano, A.V. L1 (LINE-1) retrotransposon evolution and amplification in recent human history. Mol. Biol. Evol. 17, 915–928 (2000).

    CAS  Article  Google Scholar 

  24. Kazazian, H.H. Jr. An estimated frequency of endogenous insertional mutations in humans. Nature Genet. 22, 130 (1999).

  25. Li, X. et al. Frequency of recent retrotransposition events in the human factor IX gene. Hum. Mutat. 17, 511–519 (2001).

    CAS  Article  Google Scholar 

  26. Jensen, S., Gassama, M.P. & Heidmann, T. Taming of transposable elements by homology-dependent gene silencing. Nature Genet. 21, 209–212 (1999).

    CAS  Article  Google Scholar 

  27. Hohjoh, H. & Singer, M.F. Cytoplasmic ribonucleoprotein complexes containing human LINE-1 protein and RNA. EMBO J. 15, 630–639 (1996).

    CAS  Article  Google Scholar 

  28. Kolosha, V.O. & Martin, S.L. In vitro properties of the first ORF protein from mouse LINE-1 support its role in ribonucleoprotein particle formation during retrotransposition. Proc. Natl Acad. Sci. USA 94, 10155–10160 (1997).

    CAS  Article  Google Scholar 

  29. Mathias, S.L., Scott, A.F., Kazazian, H.H., Boeke, J.D. & Gabriel, A. Reverse transcriptase encoded by a human transposable element. Science 254, 1808–1810 (1991).

    CAS  Article  Google Scholar 

  30. Fanning, T. & Singer, M. The line-1 DNA-sequences in 4 mammalian orders predicts proteins that conserve homologies to retrovirus proteins. Nucleic Acids Res. 15, 2251–2260 (1987).

    CAS  Article  Google Scholar 

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Acknowledgements

We thank A. Farley, K.-S. Kim and M.C. Cha for technical assistance; the Transgenic and DNA Sequencing Cores of the University of Pennsylvania for generation of transgenic mouse lines and for DNA sequences, respectively; and J. Moran and B. Brouha for critical reading of the manuscript. E.M.O. was supported by a Howard Hughes Predoctoral Fellowship, and H.H.K. was supported by grants from the US National Institutes of Health.

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Affiliations

  1. Department of Genetics and University of Pennsylvania School of Medicine, 475 Clinical Research Bldg., 415 Curie Blvd., Philadelphia, 19104, Pennsylvania, USA

    Eric M. Ostertag, Ralph J. DeBerardinis, John L. Goodier, Yue Zhang, Nuo Yang & Haig H. Kazazian Jr.

  2. Center for Research on Reproduction and Women's Health, University of Pennsylvania School of Medicine, 475 Clinical Research Bldg., 415 Curie Blvd., Philadelphia, 19104, Pennsylvania, USA

    George L. Gerton

Authors
  1. Eric M. Ostertag
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  2. Ralph J. DeBerardinis
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  3. John L. Goodier
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  4. Yue Zhang
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  5. Nuo Yang
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  6. George L. Gerton
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  7. Haig H. Kazazian Jr.
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Correspondence to Haig H. Kazazian Jr..

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Ostertag, E., DeBerardinis, R., Goodier, J. et al. A mouse model of human L1 retrotransposition. Nat Genet 32, 655–660 (2002). https://doi.org/10.1038/ng1022

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