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Lentivirus Gene Engineering Protocols pp 149–160Cite as

Manipulating the Cell Differentiation Through Lentiviral Vectors

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 614))

Abstract

The manipulation of cell differentiation is important to create new sources for the treatment of degenerative diseases or solve cell depletion after aggressive therapy against cancer. In this chapter, the use of a tissue-specific promoter lentiviral vector to obtain a myocardial pure lineage from murine embryonic stem cells (mES) is described in detail. Since the cardiac isoform of troponin I gene product is not expressed in skeletal or other muscle types, short mouse cardiac troponin proximal promoter is used to drive reporter genes. Cells are infected simultaneously with two lentiviral vectors, the first expressing EGFP to monitor the transduction efficiency, and the other expressing a puromycin resistance gene to select the specific cells of interest. This technical approach describes a method to obtain a pure cardiomyocyte population and can be applied to other lineages of interest.

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Acknowledgments

The authors thank Antonio Addario e Giuseppe Loreto for their professional technical support. The authors also thank Antonia Follenzi and Luigi Naldini for providing lentiviral vectors. This work was supported by the Italian Health Ministry and Istituto Superiore di Sanità.

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Authors and Affiliations

  1. Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore Sanità, Rome, Italy

    Valeria Coppola, Cesare Galli, Maria Musumeci & Désirée Bonci

Authors
  1. Valeria Coppola
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  2. Cesare Galli
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  3. Maria Musumeci
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  4. Désirée Bonci
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Editor information

Editors and Affiliations

  1. Div. Patogenesi dei Retrovirus, Centro Nazionale AIDS, Viale Regina Elena 299, Roma, 00161, Italy

    Maurizio Federico

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© 2010 Humana Press, a part of Springer Science+Business Media, LLC

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Coppola, V., Galli, C., Musumeci, M., Bonci, D. (2010). Manipulating the Cell Differentiation Through Lentiviral Vectors. In: Federico, M. (eds) Lentivirus Gene Engineering Protocols. Methods in Molecular Biology, vol 614. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-533-0_10

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  • DOI: https://doi.org/10.1007/978-1-60761-533-0_10

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-532-3

  • Online ISBN: 978-1-60761-533-0

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