Journal
NUCLEIC ACIDS RESEARCH
Volume 38, Issue 22, Pages -Publisher
OXFORD UNIV PRESS
DOI: 10.1093/nar/gkq860
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Funding
- Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan [20700368, 16651102]
- Tokai University School of Medicine
- Tokai University Educational System General Research Organization
- Institute of Medical Sciences in Tokai University
- Grants-in-Aid for Scientific Research [20700368, 16651102] Funding Source: KAKEN
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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.
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