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Using the 2A Protein Coexpression System: Multicistronic 2A Vectors Expressing Gene(s) of Interest and Reporter Proteins

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Reporter Gene Assays

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1755))

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Abstract

To date, a huge range of different proteins—many with cotranslational and posttranslational subcellular localization signals—have been coexpressed together with various reporter proteins in vitro and in vivo using 2A peptides. The pros and cons of 2A co-expression technology are considered below, followed by a simple example of a “how to” protocol to concatenate multiple genes of interest, together with a reporter gene, into a single gene linked via 2As for easy identification or selection of transduced cells.

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Acknowledgments

The authors gratefully acknowledge the long-term support of our research by the UK Biotechnology and Biological Sciences Research Council (BBSRC) and the Wellcome Trust. The University of St Andrews is a charity registered in Scotland, no. SCO13532.

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Authors and Affiliations

  1. Biomedical Sciences Research Complex, School of Biology, University of St Andrews, Fife, Scotland, UK

    Garry A. Luke & Martin D. Ryan

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  1. Garry A. Luke
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  2. Martin D. Ryan
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Correspondence to Garry A. Luke .

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Editors and Affiliations

  1. Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, California, USA

    Robert Damoiseaux

  2. Pfizer, Inc., Cambridge, Massachusetts, USA

    Samuel Hasson

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Luke, G.A., Ryan, M.D. (2018). Using the 2A Protein Coexpression System: Multicistronic 2A Vectors Expressing Gene(s) of Interest and Reporter Proteins. In: Damoiseaux, R., Hasson, S. (eds) Reporter Gene Assays. Methods in Molecular Biology, vol 1755. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7724-6_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7724-6_3

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