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Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines

Gene Therapy volume 21, pages 645–652 (2014)Cite this article

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Abstract

The use of DNA to deliver vaccine antigens offers many advantages, including ease of manufacture and cost. However, most DNA vaccines are plasmids and must be grown in bacterial culture, necessitating elements that are either unnecessary for effective gene delivery (for example, bacterial origins of replication) or undesirable (for example, antibiotic resistance genes). Removing these elements may improve the safety profile of DNA for the delivery of vaccines. Here, we describe a novel, double-stranded, linear DNA construct produced by an enzymatic process that solely encodes an antigen expression cassette, comprising antigen, promoter, polyA tail and telomeric ends. We compared these constructs (called ‘Doggybones’ because of their shape) with conventional plasmid DNA. Using luciferase-expressing constructs, we demonstrated that expression levels were equivalent between Doggybones and plasmids both in vitro and in vivo. When mice were immunized with DNA constructs expressing the HIV envelope protein gp140, equivalent humoral and cellular responses were induced. Immunizations with either construct type expressing hemagluttinin were protective against H1N1 influenza challenge. This is the first example of an effective DNA vaccine, which can be produced on a large scale by enzymatic processes.

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Acknowledgements

We would like to thank Qin Hu (St George’s University of London) for the MWS2 gp140 gene sequence, Ruth Elderfield and Wendy Barclay (Imperial College London) for the influenza hemagluttinin gene sequence and influenza virus and Paul Rothwell, Jon Extance, Karen Oliver and Kinga Karbowniczek (Touchlight Genetics Ltd.) for the generation of the Doggybone material. This work was funded in part by Touchlight Genetics Ltd. and the Exgenomes project under the Seventh Framework program for funding. EK is funded by an MRC CASE studentship MR/J006548/1. We are indebted to Dormeur Investment Service Ltd. for funding of equipment used in this project.

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Author notes

  1. A A Walters

    Present address: 3Current address: The Jenner Institute, University of Oxford, Oxford OX3 7DQ, UK.,

Authors and Affiliations

  1. Department of Medicine, Mucosal Infection & Immunity Group, Section of Infectious Diseases, Imperial College London, St Mary’s Campus, London, UK

    A A Walters, E Kinnear, R J Shattock, J U McDonald & J S Tregoning

  2. Touchlight Genetics Ltd., Leatherhead Food Research Institute, Leatherhead, Surrey, UK

    L J Caproni & N Porter

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  1. A A Walters
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  2. E Kinnear
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  3. R J Shattock
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  7. J S Tregoning
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Corresponding author

Correspondence to J S Tregoning.

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Competing interests

The study was funded in part by Touchlight Genetics, who were involved in all stages of the study conduct and analysis. Dr Caproni and Dr Porter are employees of Touchlight Genetics. The corresponding author had final responsibility to submit for publication.

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Walters, A., Kinnear, E., Shattock, R. et al. Comparative analysis of enzymatically produced novel linear DNA constructs with plasmids for use as DNA vaccines. Gene Ther 21, 645–652 (2014). https://doi.org/10.1038/gt.2014.37

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