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Codon-Optimized Genes that Enable Increased Heterologous Expression in Mammalian Cells and Elicit Efficient Immune Responses in Mice after Vaccination of Naked DNA

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Molecular Diagnosis of Infectious Diseases

Part of the book series: Methods in Molecular Medicine™ ((MIMM,volume 94))

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Abstract

Many of the problems related with mammalian gene expression, such as low translation efficiency and mRNA halflife, can be solved by means of a rational gene design, based on modern bioinformatics, followed by the de novo generation of a synthetic gene. Moreover, high expression rates and prolonged mRNA stability are not only crucial for heterologous mammalian expression, but, in particular, are important for the generation of effective DNA vaccines. In this chapter we show that an optimized synthetic gene encoding the HIV-1 Pr55gag outperforms wild-type gene driven expression by several orders of magnitude. RNA analysis revealed that this positive effect was mostly due to increased mRNA stability of the optimized transcripts. Moreover, mice vaccinated with the optimized gag gene elicited a much stronger immune response against Pr55gag than the control groups immunized with the respective wild-type gene.

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Author information

Authors and Affiliations

  1. Geneart, Regensburg, Germany

    Marcus Graf

  2. Institute for Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany

    Ludwig Deml & Ralf Wagner

Authors
  1. Marcus Graf
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  2. Ludwig Deml
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  3. Ralf Wagner
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Editors and Affiliations

  1. Accenture GmbH, Munich, Germany

    Jochen Decler

  2. Institute for Medical Microbiology and Hygiene, University of Regensburg, Regenburg, Germany

    Udo Reischl

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© 2004 Humana Press Inc.

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Graf, M., Deml, L., Wagner, R. (2004). Codon-Optimized Genes that Enable Increased Heterologous Expression in Mammalian Cells and Elicit Efficient Immune Responses in Mice after Vaccination of Naked DNA. In: Decler, J., Reischl, U. (eds) Molecular Diagnosis of Infectious Diseases. Methods in Molecular Medicine™, vol 94. Humana Press. https://doi.org/10.1385/1-59259-679-7:197

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  • DOI: https://doi.org/10.1385/1-59259-679-7:197

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-221-6

  • Online ISBN: 978-1-59259-679-9

  • eBook Packages: Springer Protocols

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