Proceedings of the National Academy of Sciences · 2012 · 59 citations · 34 references
Veterinary MedicineBiotechnologyGeneticsAssisted Reproductive TechnologiesEmbryo TransferTransgenic Animal ModelsEmbryo ManipulationEmbryo Transfer TechnologyIntracytoplasmic Sperm InjectionPronuclear MicroinjectionTransgenesis StrategiesTransgenic Animal TechnologyEmbryo MicroinjectionIntracytoplasmic Sperm Injection FertilizationMicroinjectionTransgene ExpressionTransgene IntegrationTransgenic AnimalsTransgenic MiceTransgenic Mouse Models
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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Production of transgenic rabbits, sheep and pigs by microinjection
Robert E. Hammer, V. G. Pursel, Caird E. Rexroad et al. · Nature · 1985 · 1.1K citations