NDLI: Site-specific integration of transgene targeting an endogenous lox-like site in early mouse embryos

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Site-specific integration of transgene targeting an endogenous lox-like site in early mouse embryos

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Journal of Applied Genetics : Volume 51

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Journal of Applied Genetics : Volume 48

Journal of Applied Genetics : Volume 47

Site-specific integration of transgene targeting an endogenous lox-like site in early mouse embryos

Content Provider	Springer Nature Link
Author	Ito, Masanori Yamanouchi, Keitaro Naito, Kunihiko Calos, Michele P. Tojo, Hideaki
Copyright Year	2010
Abstract	Functional lox-like sequences have been identified within the yeast and mammalian genome. These hetero-specific lox sites also allow Cre recombinase to specifically target efficient integration of exogenous DNA into the endogenous pseudo-lox (ψlox) sequences that occur naturally in the host genome using a Cre/loxP integrative recombination system. We investigated whether the Cre/ψlox system is useful for site-specific integration of transgenes and for improving the production efficiency of transgenic animals. This is the first report on Cre-mediated integrative recombination targeting an endogenous lox-like sequence termed pseudo-loxm5 (ψloxm5) in early mouse embryos. We characterized the Cre/ψloxm5 system in embryonic environment. Cre-expressing plasmid and a transgene (CMV/LacZ gene) flanked by ψloxm5 and ψloxcorem5 sites were co-microinjected into the pronucleus of fertilized mouse oocytes. The injected eggs were transferred into foster mothers, and the recombination products were investigated. The results show that the ψloxm5 site is an active substrate for Cre-mediated recombination in the mouse embryonic environment. The transgenesis efficiency was up to 27% (6/22). The site-specific integration of the transgene into the endogenous ψloxm5 site was found in 50 % of the transgenic pups. Our findings demonstrated that the Cre/ψloxm5 integrative recombination system is an efficient and simple strategy for targeting an endogenous lox-like site in mammalian embryos.
Starting Page	89
Ending Page	94
Page Count	6
File Format	PDF
ISSN	12341983
Journal	Journal of Applied Genetics
Volume Number	52
Issue Number	1
e-ISSN	21903883
Language	English
Publisher	Springer-Verlag
Publisher Date	2010-11-26
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Cre Microinjection Pseudo-lox Site-specific integration Transgenic animal Plant Genetics & Genomics Microbial Genetics and Genomics Human Genetics Animal Genetics and Genomics Life Sciences
Content Type	Text
Resource Type	Article
Subject	Genetics Medicine

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