Helper-independent piggyBac plasmids for gene delivery approaches: Strategies for avoiding potential genotoxic effects

Johann Urschitz, Miyuri Kawasumi, Jesse Owens, Kazuto Morozumi, Hideaki Yamashiro, Ilko Stoytchev, Joel Marh, James A. Dee, Kris Kawamoto, Craig J. Coates, Joseph M. Kaminski, Pawel Pelczar, Ryuzo Yanagimachi, Stefan Moisyadi

Research output: Contribution to journalArticle

44 Scopus citations

Abstract

Efficient integration of functional genes is an essential prerequisite for successful gene delivery such as cell transfection, animal transgenesis, and gene therapy. Gene delivery strategies based on viral vectors are currently the most efficient. However, limited cargo capacity, host immune response, and the risk of insertional mutagenesis are limiting factors and of concern. Recently, several groups have used transposon-based approaches to deliver genes to a variety of cells. The piggyBac (pB) transposase in particular has been shown to be well suited for cell transfection and gene therapy approaches because of its flexibility for molecular modification, large cargo capacity, and high transposition activity. However, safety considerations regarding transposase gene insertions into host genomes have rarely been addressed. Here we report our results on engineering helper-independent pB plasmids. The single-plasmid gene delivery system carries both the piggyBac transposase (pBt) expression cassette as well as the transposon cargo flankedby terminal repeat element sequences. Improvements to the helper-independent structure were achieved by developing new plasmids in which the pBt gene is rendered inactive after excision of the transposon from the plasmid. As a consequence, potentially negative effects that may develop by the persistence of an active pBt gene posttransposition are eliminated. The results presented herein demonstrate that our helper-independent plasmids represent an important step in the development of safe and efficient gene delivery methods that should prove valuable in gene therapy and transgenic approaches.

Original languageEnglish (US)
Pages (from-to)8117-8122
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume107
Issue number18
DOIs
StatePublished - May 4 2010

Keywords

  • Gene therapy
  • Transfection
  • Transgenesis
  • Transposase
  • Transposon

ASJC Scopus subject areas

  • General

Fingerprint Dive into the research topics of 'Helper-independent piggyBac plasmids for gene delivery approaches: Strategies for avoiding potential genotoxic effects'. Together they form a unique fingerprint.

  • Cite this

    Urschitz, J., Kawasumi, M., Owens, J., Morozumi, K., Yamashiro, H., Stoytchev, I., Marh, J., Dee, J. A., Kawamoto, K., Coates, C. J., Kaminski, J. M., Pelczar, P., Yanagimachi, R., & Moisyadi, S. (2010). Helper-independent piggyBac plasmids for gene delivery approaches: Strategies for avoiding potential genotoxic effects. Proceedings of the National Academy of Sciences of the United States of America, 107(18), 8117-8122. https://doi.org/10.1073/pnas.1003674107