title: PITT: pronuclear injection-based targeted transgenesis, a reliable transgene expression method in mice.
abstract: Transgenic (Tg) mice have been extensively used as valuable tools for analyses of gene function and have also served as models for many human diseases. Typically, a transgenic mouse is created by microinjection of DNA into pronuclei in which the DNA gets integrated at random locations in the genome. Frequently however, the random integration of multiple copies of a transgene results in transgene silencing, probably because of a positional effect and/or repeat-induced gene silencing. The transgene silencing issue has been overcome by single-copy transgene integration into a predetermined locus through ES cell-mediated transgenesis, despite it being expensive and more time-consuming compared with pronuclear injection (PI)-mediated transgenesis. Recently, several groups have reported novel approaches that employ PI for targeted transgenesis. They are based on site-specific recombination catalyzed by a recombinase or an integrase or homologous recombination enhanced by a zinc-finger nuclease via PI. These next-generation transgenesis methods, which we termed as PI-based Targeted Transgenesis (PITT), are more convenient and faster than ES cell-based transgenesis. Furthermore, the Tg mice generated by these newer methods contain a single-copy transgene and exhibit reliable expression of the transgene. The objective of this review is to present the recent progress in mouse targeted transgenesis.
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.
authors.forename: Masato
authors.initials: M
authors.lastname: Ohtsuka
authors.forename: Hiromi
authors.initials: H
authors.lastname: Miura
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sato
authors.forename: Minoru
authors.initials: M
authors.lastname: Kimura
authors.forename: Hidetoshi
authors.initials: H
authors.lastname: Inoko
authors.forename: Channabasavaiah B
authors.initials: CB
authors.lastname: Gurumurthy
mesh_terms: D000818:Animals; D004254:DNA Nucleotidyltransferases / Q000378:metabolism; D015870:Gene Expression; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D019426:Integrases / Q000378:metabolism; D051379:Mice / Q000235:genetics*; D008822:Mice, Transgenic; D008845:Microinjections; D053652:Nuclear Transfer Techniques; D011995:Recombination, Genetic; D012260:Ribonucleases / Q000378:metabolism; D019076:Transgenes; D016335:Zinc Fingers
chemical_list: D004254:DNA Nucleotidyltransferases; D019426:Integrases; C077570:Site-specific recombinase; D012260:Ribonucleases