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

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.

SUBMITTER: Marh J

PROVIDER: S-EPMC3511126 | biostudies-literature | 2012 Nov

REPOSITORIES: biostudies-literature

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

Marh Joel J Stoytcheva Zoia Z Urschitz Johann J Sugawara Atsushi A Yamashiro Hideaki H Owens Jesse B JB Stoytchev Ilko I Pelczar Pawel P Yanagimachi Ryuzo R Moisyadi Stefan S

Proceedings of the National Academy of Sciences of the United States of America 20121023 47

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 m ...[more]

PMID: 23093669

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