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

doi:10.1073/pnas.1216473109

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

HOSP - Obstetrics, Gynecology and Urology

Research output: Contribution to journal › Article › peer-review

38 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 language	English
Pages (from-to)	19184-19189
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	109
Issue number	47
DOIs	https://doi.org/10.1073/pnas.1216473109
Publication status	Published - 2012 Nov 20

Keywords

Genetically modified
Transgene

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10.1073/pnas.1216473109

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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

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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.",

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Marh, J, Stoytcheva, Z, Urschitz, J, Sugawara, A, Yamashiro, H, Owens, JB, 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, vol. 109, no. 47, pp. 19184-19189. https://doi.org/10.1073/pnas.1216473109

TY - JOUR

T1 - Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis

AU - Marh, Joel

AU - Stoytcheva, Zoia

AU - Urschitz, Johann

AU - Sugawara, Atsushi

AU - Yamashiro, Hideaki

AU - Owens, Jesse B.

AU - Stoytchev, Ilko

AU - Pelczar, Pawel

AU - Yanagimachi, Ryuzo

AU - Moisyadi, Stefan

PY - 2012/11/20

Y1 - 2012/11/20

N2 - 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.

AB - 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.

KW - Genetically modified

KW - Transgene

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JF - Proceedings of the National Academy of Sciences of the United States of America

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ER -

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

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