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APOBEC3A catabolism of electroporated plasmid DNA in mouse muscle

Gene Therapy volume 22, pages 96–103 (2015)Cite this article

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Abstract

The mouse is widely used as a model for DNA therapy and vaccination even though the efficiency of DNA delivery in higher mammals and humans is much less. The human APOBEC3 (A3) enzymes impact viral genomes by cytidine deamination, which introduces multiple uridine residues into single-stranded DNA, a process known as genetic editing. This initiates rapid DNA catabolism via a uracil DNA glycosylase dependent pathway. In tissue culture, A3A, A3C and A3B can hyperedit transfected plasmid DNA. We explored plasmid catabolism in vivo initiated by A3A, the most efficient of the human enzymes and one that is functionally conserved across most mammals. As rodents do not encode an A3A enzyme, it was possible to explore DNA degradation in the mouse model. Human A3A genetically edits co-electroporated luciferase plasmid DNA in mouse skeletal muscle that initiates DNA degradation resulting in approximately fourfold decrease in bioluminescence. Part of the degradation occurs in the nucleus as indicated by complex hyperedited DNA molecules. As human A3A is strongly upregulated by interferon α and DNA sensing pathways, it is a strong candidate enzyme for restricting plasmid DNA in higher mammals.

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Acknowledgements

This work was supported by grants from Institut Pasteur and Invectys. Marie Escande for technical support in tissue dissection.

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Authors and Affiliations

  1. Invectys, Pasteur BioTop, Institut Pasteur, CNRS URA 3015, France

    A Kostrzak, P L Demoyen & S Wain-Hobson

  2. Molecular Retrovirology Unit, Institut Pasteur, CNRS URA 3015, France

    M Henry, S Wain-Hobson & J-P Vartanian

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  1. A Kostrzak
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Correspondence to J-P Vartanian.

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PLD and SWH are co-founders of Invectys, a biotech devoted to therapeutic cancer vaccination. The remaining authors declare no conflict of interest.

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Kostrzak, A., Henry, M., Demoyen, P. et al. APOBEC3A catabolism of electroporated plasmid DNA in mouse muscle. Gene Ther 22, 96–103 (2015). https://doi.org/10.1038/gt.2014.88

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