title: Development of pronuclear injection-based targeted transgenesis in mice through Cre-loxP site-specific recombination.
abstract: Microinjection of DNA into the pronuclei of zygotes is the simplest and most widely used method for generating transgenic (Tg) mice. However, it is always associated with random integration of multiple copies of the transgene, resulting in unstable, low, or no transgene expression due to positional effects and/or repeat-induced gene silencing. In addition, random integration sometimes disrupts an endogenous gene that can affect the phenotypes of Tg mice. Our recently developed pronuclear injection-based targeted transgenesis (PITT) method enables the integration of a single-copy transgene into a predetermined genomic locus through Cre-loxP site-specific recombination. The PITT method enables stable and reliable transgene expression in Tg mice and is also applicable for generating knockdown mice. Therefore, the PITT method could represent next-generation transgenesis that overcomes the pitfalls of conventional transgenesis.
authors: Division of Basic Medical Science and Molecular Medicine, Department of Molecular Life Science, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa, 259-1193, Japan, masato@is.icc.u-tokai.ac.jp.
authors: Masato
authors: M
authors: Ohtsuka
mesh_terms: D000818:Animals; D001287:Attachment Sites, Microbiological / Q000235:genetics*; D004624:Embryo Transfer; D005260:Female; D005307:Fertilization in Vitro; D018390:Gene Targeting / Q000379:methods*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D060005:Genotyping Techniques; D019426:Integrases / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008845:Microinjections; D012333:RNA, Messenger / Q000235:genetics; D011995:Recombination, Genetic; D019076:Transgenes / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; C045073:Cre recombinase; D019426:Integrases