L1 retrotransposition in human neural progenitor cells

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Abstract

Long interspersed element 1 (LINE-1 or L1) retrotransposons have markedly affected the human genome. L1s must retrotranspose in the germ line or during early development to ensure their evolutionary success, yet the extent to which this process affects somatic cells is poorly understood. We previously demonstrated that engineered human L1s can retrotranspose in adult rat hippocampus progenitor cells in vitro and in the mouse brain in vivo1. Here we demonstrate that neural progenitor cells isolated from human fetal brain and derived from human embryonic stem cells support the retrotransposition of engineered human L1s in vitro. Furthermore, we developed a quantitative multiplex polymerase chain reaction that detected an increase in the copy number of endogenous L1s in the hippocampus, and in several regions of adult human brains, when compared to the copy number of endogenous L1s in heart or liver genomic DNAs from the same donor. These data suggest that de novo L1 retrotransposition events may occur in the human brain and, in principle, have the potential to contribute to individual somatic mosaicism.

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Figure 1: L1 retrotransposition in hCNS-SCns.
Figure 2: L1 retrotransposition in hESC-derived NPCs.
Figure 3: Methylation analysis and ChIP for the endogenous human L1 5′ UTR.
Figure 4: Multiplex quantitative PCR analyses of L1 copy number in human tissues.

Change history

  • 27 August 2009

    The position of the 'DAPI' label on Fig. 1f was altered on 27 August 2009.

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Acknowledgements

We thank J. Simon for excellent schematic drawings, M. L. Gage, J. Kim and H. Kopera for editorial comments, B. Miller and R. Keithley for cell culture assistance, C. T. Carson for hESC advice, D. Chambers and J. Barrie for flow cytometry assistance, L. Randolph-Moore for molecular advice, B. Aimone for statistics advice, T. Liang for microarray assistance, and Y. Lineu and J. Mosher for helpful comments. We also thank T. Fanning and M. Klymkowsky for the ORF1 protein and SOX3 antibodies, respectively. F.H.G. and N.G.C. are supported by the Picower Foundation, G. Harold and Leila Y. Mathers Charitable Foundation, Lookout Fund (MH082070), and the California Institute for Regenerative Medicine (CIRM). J.L.G.-P. is supported by Plan Estabilizacion Grupos SNS ENCYT 2015 (EMER07/56, Instituto de Salud Carlos III, Spain) and through the IRG-FP7-PEOPLE-2007 Marie Curie program. K.S.O. was supported by grants GM069985 and NS048187 from the National Institutes of Health (NIH). J.V.M. was supported by grants GM082970 and GM069985 from the NIH and by the Howard Hughes Medical Institute. Work in the laboratories of K.S.O. and J.V.M. only used NIH-approved stem cell lines.

Author Contributions N.G.C. and F.H.G. directed the project. J.V.M. and J.L.G.-P. directed aspects of the project conducted at Michigan. N.G.C., J.L.G.-P., J.V.M. and F.H.G. designed experiments and drafted the manuscript. N.G.C., F.H.G., J.L.G-P. and G.E.P. performed the experiments. G.W.Y. and M.T.L. carried out bioinformatics data analysis. Y.M. performed electrophysiology experiments. M.M. and K.S.O. provided hESC culture and NPC differentiation assistance. All authors commented on or contributed to the current manuscript.

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Correspondence to Fred H. Gage.

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Coufal, N., Garcia-Perez, J., Peng, G. et al. L1 retrotransposition in human neural progenitor cells. Nature 460, 1127–1131 (2009) doi:10.1038/nature08248

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