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Analysis of LINE-1 Expression in Human Pluripotent Cells

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Human Embryonic Stem Cells Handbook

Part of the book series: Methods in Molecular Biology ((MIMB,volume 873))

Abstract

Half of the human genome is composed of repeated DNA, and some types are mobile within our genome (transposons and retrotransposons). Despite their abundance, only a small fraction of them are currently active in our genome (Long Interspersed Element-1 (LINE-1), Alu, and SVA elements). LINE-1 or L1 elements are a family of active non-LTR retrotransposons, the ongoing mobilization of which still impacts our genome. As selfish DNA elements, L1 activity is more prominent in early human development, where new insertions would be transmitted to the progeny. Here, we describe the conventional methods aimed to determine the expression level of LINE-1 elements in pluripotent human cells.

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Acknowledgments

M.M.-L. and M.G.-C. contributed equally to this chapter. We thank Gael Cristofari (CNRS, France) for the anti-L1.3-ORF1p antibody. J.L.G.-P. lab is supported by a Marie Curie IRG action (FP7-PEOPLE-2007-4-3-IRG), Instituto de Salud Carlos III/FEDER, Spain (EMER07/056, CP07/00065, and FIS/FEDER PI08171), and Junta de Andalucia, Spain (CICE/FEDER P09-CTS-4980 and PeS-FEDER PI-002). M.M.-L. and S.M are supported by CICE P09-CTS-4980 and CICE P08-CTS-3678 from Junta de Andalucia, Spain. We thank the valuable input from colleagues at our institution. We also thank the contribution of the laboratories of Fred H. Gage (Salk Institute, USA) and John V. Moran (Howard Hughes Medical Institute, University of Michigan, USA) for their work in many of the techniques included in this chapter.

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Authors and Affiliations

  1. Andalusian Stem Cell Bank, Consejeria de Salud Junta de Andalucia, Seville, Spain

    Martin Muñoz-Lopez, Marta Garcia-Cañadas, Angela Macia, Santiago Morell & Jose L. Garcia-Perez

  2. Center for Biomedical Research, University of Granada, Armilla, Granada, Spain

    Martin Muñoz-Lopez, Marta Garcia-Cañadas, Angela Macia, Santiago Morell & Jose L. Garcia-Perez

Authors
  1. Martin Muñoz-Lopez
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  2. Marta Garcia-Cañadas
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  3. Angela Macia
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  4. Santiago Morell
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  5. Jose L. Garcia-Perez
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Corresponding author

Correspondence to Jose L. Garcia-Perez .

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Editors and Affiliations

  1. Sprott Centre for Stem Cell Research, Regenerative Medicine Program, Ottawa Hospital Research Institute, Smyth Road 501, Ottawa, K1Y 8L6, Canada

    Kursad Turksen

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Muñoz-Lopez, M., Garcia-Cañadas, M., Macia, A., Morell, S., Garcia-Perez, J.L. (2012). Analysis of LINE-1 Expression in Human Pluripotent Cells. In: Turksen, K. (eds) Human Embryonic Stem Cells Handbook. Methods in Molecular Biology, vol 873. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-794-1_7

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  • DOI: https://doi.org/10.1007/978-1-61779-794-1_7

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-793-4

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