title: 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.
authors.affiliation: Department of Anatomy, John A. Burns School of Medicine, University of Hawaii, Honolulu, HI 96822, USA.
authors.forename: Joel
authors.initials: J
authors.lastname: Marh
authors.forename: Zoia
authors.initials: Z
authors.lastname: Stoytcheva
authors.forename: Johann
authors.initials: J
authors.lastname: Urschitz
authors.forename: Atsushi
authors.initials: A
authors.lastname: Sugawara
authors.forename: Hideaki
authors.initials: H
authors.lastname: Yamashiro
authors.forename: Jesse B
authors.initials: JB
authors.lastname: Owens
authors.forename: Ilko
authors.initials: I
authors.lastname: Stoytchev
authors.forename: Pawel
authors.initials: P
authors.lastname: Pelczar
authors.forename: Ryuzo
authors.initials: R
authors.lastname: Yanagimachi
authors.forename: Stefan
authors.initials: S
authors.lastname: Moisyadi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002467:Cell Nucleus / Q000378:metabolism*; D002478:Cells, Cultured; D003433:Crosses, Genetic; D004251:DNA Transposable Elements / Q000235:genetics*; D004622:Embryo, Mammalian / Q000378:metabolism; D005260:Female; D018014:Gene Transfer Techniques; D016678:Genome / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008845:Microinjections / Q000379:methods*; D016254:Mutagenesis, Insertional / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D020554:Sperm Injections, Intracytoplasmic; D013997:Time Factors; D014162:Transfection; D019076:Transgenes / Q000235:genetics; D019895:Transposases / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D004251:DNA Transposable Elements; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D019895:Transposases