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In Vitro-Transcribed (IVT)-mRNA CAR Therapy Development

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Chimeric Antigen Receptor T Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2086))

Abstract

Chimeric antigen receptor (CAR) cancer immunotherapy uses autologous immune system’s cells, genetically modified, to reinforce the immune system against cancer cells. Genetic modification is usually mediated via viral transfection, despite the risk of insertional oncogenesis and off target side effects. In vitro-transcribed (IVT)-mRNA-mediated transfection could contribute to a much safer CAR therapy, since IVT-mRNA leaves no ultimate genetic residue in recipient cells. In this chapter, the IVT-mRNA generation procedure is described, from the selection of the target of the CAR T-cells, the cloning of the template for the in vitro transcription and the development of several chemical modifications for optimizing the structure and thus the stability of the produced CAR IVT-mRNA molecules. Among various transfection methods to efficiently express the CAR molecule on T-cells’ surface, the electroporation and the cationic-lipid mediated transfection of the CAR IVT-mRNAs are described.

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Change history

  • 03 January 2020

    This chapter was inadvertently published with one of the contributing author’s name printed as Miliotou N. Androulla, whereas it should have been A. N. Miliotou . This correction has been updated in the book.

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Authors and Affiliations

  1. Laboratory of Pharmacology, School of Pharmacy, Faculty of Health Sciences, Aristotle University of Thessaloniki, Thessaloniki, Macedonia, Greece

    Androulla N. Miliotou & Lefkothea C. Papadopoulou

Authors
  1. Androulla N. Miliotou
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  2. Lefkothea C. Papadopoulou
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Corresponding author

Correspondence to Lefkothea C. Papadopoulou .

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Editors and Affiliations

  1. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Kamilla Swiech

  2. School of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Kelen Cristina Ribeiro Malmegrim

  3. Center for Cell-Based Therapy CTC Regional Blood Center of Ribeirão Preto, University of São Paulo, Ribeirão Preto, São Paulo, Brazil

    Virgínia Picanço-Castro

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Miliotou, A.N., Papadopoulou, L.C. (2020). In Vitro-Transcribed (IVT)-mRNA CAR Therapy Development. In: Swiech, K., Malmegrim, K., Picanço-Castro, V. (eds) Chimeric Antigen Receptor T Cells. Methods in Molecular Biology, vol 2086. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0146-4_7

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  • DOI: https://doi.org/10.1007/978-1-0716-0146-4_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0145-7

  • Online ISBN: 978-1-0716-0146-4

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