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Stable Overexpression of miRNAs in Bone Marrow-Derived Murine Mast Cells Using Lentiviral Expression Vectors

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MicroRNAs and the Immune System

Part of the book series: Methods in Molecular Biology ((MIMB,volume 667))

Abstract

MicroRNAs (miRNAs) constitute a class of molecules regulating gene expression in many different cell types, including cells of the mammalian immune system. Indeed, changes in miRNA expression patterns have been implicated in various physiological and pathological processes. Mast cells (MCs) are hematopoietic cells that originate in the bone marrow and migrate into the tissues, where they mature and reside. They have an important immunoregulatory and effector role in IgE-associated allergic disorders, as well as in certain innate and adaptive immune responses. An effective way to explore the functions of miRNAs in murine MCs includes the modification of miRNA expression in primary bone marrow-derived mast cells (BMMCs), followed by the analysis of the phenotypic consequences of such perturbation. In this chapter, we describe how to differentiate BMMCs and transduce them with lentiviruses. As an example, we expressed miR-221 and miR-222, which showed stable expression in BMMCs and acted as post-transcriptional regulators of c-Kit expression.

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Acknowledgements

We would like to thank Dr. Thomas Pertel and Dr. Desiree Bonci for the pAPM and TWEEN lentiviral vectors, respectively. RJM is the recipient of a pre-doctoral fellowship from the San Raffaele University, Milan, Italy. This work is supported in part by a Ceresio Foundation fellowship and a Swiss National Science Foundation grant to SM.

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Authors and Affiliations

  1. Institute for Research in Biomedicine, Bellinzona, Switzerland

    Ramon J. Mayoral & Silvia Monticelli

Authors
  1. Ramon J. Mayoral
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  2. Silvia Monticelli
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Correspondence to Silvia Monticelli .

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Editors and Affiliations

  1. Institute for Research in Biomedicine, Via Vincenzo Vela 6, Bellinzona, CH-6500, Switzerland

    Silvia Monticelli

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Mayoral, R.J., Monticelli, S. (2010). Stable Overexpression of miRNAs in Bone Marrow-Derived Murine Mast Cells Using Lentiviral Expression Vectors. In: Monticelli, S. (eds) MicroRNAs and the Immune System. Methods in Molecular Biology, vol 667. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-811-9_14

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  • DOI: https://doi.org/10.1007/978-1-60761-811-9_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-810-2

  • Online ISBN: 978-1-60761-811-9

  • eBook Packages: Springer Protocols

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