Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis

Joel Marh, Zoia Stoytcheva, Johann Urschitz, Atsushi Sugawara, Hideaki Yamashiro, Jesse B. Owens, Ilko Stoytchev, Pawel Pelczar, Ryuzo Yanagimachi, Stefan Moisyadi

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

We have developed a unique method for mouse transgenesis. The transposase-enhanced pronuclear microinjection (PNI) technique described herein uses the hyperactive piggyBac transposase to insert a large transgene into the mouse genome. This procedure increased transgene integration efficiency by fivefold compared with conventional PNI or intracytoplasmic sperm injection-mediated transgenesis. Our data indicate that the transposase-enhanced PNI technique additionally requires fewer embryos to be microinjected than traditional methods to obtain transgenic animals. This transposase-mediated approach is also very efficient for single-cell embryo cytoplasmic injections, offering an easy-to-implement transgenesis method to the scientific community.

Original languageEnglish
Pages (from-to)19184-19189
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume109
Issue number47
DOIs
Publication statusPublished - 2012 Nov 20

Keywords

  • Genetically modified
  • Transgene

ASJC Scopus subject areas

  • General

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    Marh, J., Stoytcheva, Z., Urschitz, J., Sugawara, A., Yamashiro, H., Owens, J. B., Stoytchev, I., Pelczar, P., Yanagimachi, R., & Moisyadi, S. (2012). Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis. Proceedings of the National Academy of Sciences of the United States of America, 109(47), 19184-19189. https://doi.org/10.1073/pnas.1216473109