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Minimal piggyBac vectors for chromatin integration

Gene Therapy volume 21, pages 1–9 (2014)Cite this article

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Abstract

We describe novel transposon piggyBac vectors engineered to deliver transgenes as efficiently as currently available piggyBac systems, but with significantly less helper DNA co-delivered into the host genome. To generate these plasmids, we identified a previously unreported aspect of transposon biology, that the full-length terminal domains required for successful plasmid-to-chromatin transgene delivery can be removed from the transgene delivery cassette to other parts of the plasmid without significantly impairing transposition efficiency. This is achieved by including in the same plasmid, an additional helper piggyBac sequence that contains both long terminal domains, but is modified to prevent its transposition into the host genome. This design decreases the size of the required terminal domains within the delivered gene cassette of the piggyBac vector from about 1500 to just 98 base pairs. By removing these sequences from the delivered gene cassette, they are no longer incorporated into the host genome which may reduce the risk of target cell transformation.

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Acknowledgements

We thank Dr Alfred M Handler (U.S. Department of Agriculture) for providing the p3E1.2 plasmid. This work was supported in part by Grant-in-Aid awards from the Greater SouthEast Affiliate of the American Heart Association to VS (12GRNT12070291) and BF (09GRNT2260914).

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Authors and Affiliations

  1. Center for Lung Biology, University of South Alabama School of Medicine, AL, Mobile, USA

    V Solodushko & B Fouty

  2. Department of Pharmacology, University of South Alabama School of Medicine, AL, Mobile, USA

    V Solodushko & B Fouty

  3. NanoBio Corporation, Ann Arbor, MI, USA

    V Bitko

  4. Department of Internal Medicine University of South Alabama School of Medicine, AL, Mobile, USA

    B Fouty

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  1. V Solodushko
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  2. V Bitko
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Correspondence to V Solodushko.

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Solodushko, V., Bitko, V. & Fouty, B. Minimal piggyBac vectors for chromatin integration. Gene Ther 21, 1–9 (2014). https://doi.org/10.1038/gt.2013.52

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