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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]

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