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Cellular Localization of Engineered Human LINE-1 RNA and Proteins

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Transposons and Retrotransposons

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1400))

Abstract

The human LINE-1 retrotransposon has the ability to mobilize into a new genomic location through an intracellular replication cycle. Immunofluorescence and in situ hybridization experiments have been developed to detect subcellular localization of retrotransposition intermediates (i.e., ORF1p, ORF2p, and L1 mRNA). Currently, these protocols are also used to validate the interaction between retrotransposition complex components and potential cellular partners involved in L1 replication. Here, we describe in details methods for the identification of LINE-1 proteins and/or RNA in cells transfected with vectors expressing engineered human LINE-1 elements.

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Acknowledgements

We are grateful to Edouard Bertrand (CNRS, Institut de Genetique Moleculaire de Montpellier, France) for the gift of plasmids, probes, and sharing protocols. We thank Robert Singer (Albert Einstein College of Medicine, Yeshiva University, USA) for sharing online protocols (available at http://www.singerlab.org/protocols) and John V. Moran (Department of Human Genetics, University of Michigan, USA) for sharing plasmids. We are grateful to Nicole Lautredou at Montpellier RIO Imaging, France. This work was supported by the Institut National de la Santé Et de la Recherche Médicale (INSERM), the Centre National de la Recherche Scientifique (CNRS) and the Agence Nationale de la Recherche (ANR-12-BSV6-0003, RETROGENO) [to NG], Ministère de l’Enseignement Supérieur et de la Recherche, Association pour la Recherche contre le Cancer (ARC), and Fondation Recherche Médicale (FRM) [doctoral fellowships to AJD].

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Authors and Affiliations

  1. Institute for Research on Cancer and Aging of Nice (IRCAN), CNRS (UMR 7284), INSERM (U1081), University of Nice-Sophia-Antipolis, Nice cedex 2, France

    Aurélien J. Doucet

  2. Institut de Génétique Moléculaire de Montpellier, CNRS (UMR 5535), Montpellier Cedex 5, France

    Eugénia Basyuk

  3. Institute for Regenerative Medicine and Biotherapy, INSERM (U1183), 80 Avenue Augustin Fliche, 34295, Montpellier, France

    Nicolas Gilbert

Authors
  1. Aurélien J. Doucet
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  2. Eugénia Basyuk
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  3. Nicolas Gilbert
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Correspondence to Nicolas Gilbert .

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Editors and Affiliations

  1. Institute of Genetics and Molecular Medi, Genyo (Center for Genomics and Oncologic, Granada, Spain

    Jose L. Garcia-Pérez

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Doucet, A.J., Basyuk, E., Gilbert, N. (2016). Cellular Localization of Engineered Human LINE-1 RNA and Proteins. In: Garcia-Pérez, J. (eds) Transposons and Retrotransposons. Methods in Molecular Biology, vol 1400. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3372-3_18

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  • DOI: https://doi.org/10.1007/978-1-4939-3372-3_18

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3370-9

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