Hyperactive self-inactivating piggyBac for transposase-enhanced pronuclear microinjection transgenesis
| Autor: | Jesse B. Owens, Ryuzo Yanagimachi, Atsushi Sugawara, Pawel Pelczar, Joel Marh, Johann Urschitz, Zoia Stoytcheva, Hideaki Yamashiro, Stefan Moisyadi, Ilko Stoytchev |
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| Přispěvatelé: | University of Zurich, Yanagimachi, Ryuzo |
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Male
Time Factors Microinjections Transgene Green Fluorescent Proteins Mutagenesis (molecular biology technique) Transposases 610 Medicine & health Mice Transgenic Biology Transfection Insert (molecular biology) Mice Animals Humans 10239 Institute of Laboratory Animal Science Sperm Injections Intracytoplasmic Transgenes Microinjection Transposase Cells Cultured Crosses Genetic Cell Nucleus 1000 Multidisciplinary Multidisciplinary Genome Gene Transfer Techniques Antibodies Monoclonal Embryo Biological Sciences Embryo Mammalian Molecular biology Genetically modified organism Transgenesis Mutagenesis Insertional HEK293 Cells DNA Transposable Elements 570 Life sciences biology Female Plasmids |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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