title: Pronuclear injection-based mouse targeted transgenesis for reproducible and highly efficient transgene expression.
abstract: Mouse transgenesis has proven invaluable for analysis of gene function and generation of human disease models. We describe here the development of a pronuclear injection-based targeted transgenesis (PITT) system, involving site-specific integration in fertilized eggs. The system was applied to two different genomic target loci to generate a series of transgenic lines including fluorescent mice, which reproducibly displayed strong, ubiquitous and stable transgene expression. We also demonstrated that knockdown mice could be readily generated by PITT by taking advantage of the reproducible and highly efficient expression system. The PITT system, which circumvents the problem of unpredictable and unstable transgene expression of conventional random-integration transgenic mice, reduces the time, cost and effort needed to generate transgenic mice, and is potentially applicable to both in vivo 'gain-of-function' and 'loss-of-function' studies.
authors.affiliation: Department of Molecular Life Science, Division of Basic Medical Science and Molecular Medicine, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan. masato@is.icc.u-tokai.ac.jp
authors.forename: Masato
authors.initials: M
authors.lastname: Ohtsuka
authors.forename: Sanae
authors.initials: S
authors.lastname: Ogiwara
authors.forename: Hiromi
authors.initials: H
authors.lastname: Miura
authors.forename: Akiko
authors.initials: A
authors.lastname: Mizutani
authors.forename: Takayuki
authors.initials: T
authors.lastname: Warita
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sato
authors.forename: Kenji
authors.initials: K
authors.lastname: Imai
authors.forename: Katsuto
authors.initials: K
authors.lastname: Hozumi
authors.forename: Takehito
authors.initials: T
authors.lastname: Sato
authors.forename: Masafumi
authors.initials: M
authors.lastname: Tanaka
authors.forename: Minoru
authors.initials: M
authors.lastname: Kimura
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Inoko
mesh_terms: D000818:Animals; D002467:Cell Nucleus; D053595:Embryonic Stem Cells / Q000378:metabolism; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D018390:Gene Targeting / Q000379:methods*; D007267:Injections; D019426:Integrases / Q000378:metabolism; D051379:Mice; D008822:Mice, Transgenic; D010063:Ovum / Q000378:metabolism; D011995:Recombination, Genetic; D017384:Sequence Deletion; D019076:Transgenes
chemical_list: C045073:Cre recombinase; D019426:Integrases