Takemoto, Tatsuya Fujii Memorial Institute of Medical Sciences, Tokushima University|Graduate School of Frontier Biosciences, Osaka University Tokushima University Educator and Researcher Directory KAKEN Search Researchers
Abe, Takaya Genetic Engineering Team, RIKEN Center for Life Science Technologies
Kiyonari, Hiroshi Genetic Engineering Team, RIKEN Center for Life Science Technologies|Animal Resource Development Unit, RIKEN Center for Life Science Technologies
Nakao, Kazuki Animal Resource Development Unit, RIKEN Center for Life Science Technologies|Laboratory of Animal Resources, Center for Disease Biology and Integrative Medicine, Graduate School of Medicine, The University of Tokyo
Furuta, Yasuhide Genetic Engineering Team, RIKEN Center for Life Science Technologies|Animal Resource Development Unit, RIKEN Center for Life Science Technologies
Suzuki, Hitomi Fujii Memorial Institute of Medical Sciences, Tokushima University
Takada, Shinji Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences
Fujimori, Toshihiko Genetic Engineering Team, RIKEN Center for Life Science Technologies|Division of Embryology, National Institute for Basic Biology (NIBB)
Kondoh, Hisato Graduate School of Frontier Biosciences, Osaka University|Faculty of Life Sciences, Kyoto Sangyo University
The canonical Wnt signaling pathway plays a major role in the regulation of embryogenesis and organogenesis, where signal strength-dependent cellular responses are of particular importance. To assess Wnt signal levels in individual cells, and to circumvent the integration site-dependent bias shown in previous Wnt reporter lines, we constructed a new Wnt signal reporter mouse line R26-WntVis. Heptameric TCF/LEF1 binding sequences were combined with a viral minimal promoter to confer a graded response to the reporter depending on Wnt signal strengths. The histone H2B-EGFP fusion protein was chosen as the fluorescent reporter to facilitate single-cell resolution analyses. This WntVis reporter gene was then inserted into the ROSA26 locus in an orientation opposite to that of the endogenous gene. The R26-WntVis allele was introduced into Wnt3a−/− and Wnt3avt/− mutant mouse embryos and compared with wild-type embryos to assess its performance. The R26-WntVis reporter was activated in known Wnt-dependent tissues and responded in a graded fashion to signal intensity. This analysis also indicated that the major Wnt activity early in embryogenesis switched from Wnt3 to Wnt3a around E7.5. The R26-WntVis mouse line will be widely useful for the study of Wnt signal-dependent processes.
Genes to Cells
Copyright © 2016 The Authors.Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non- commercial and no modifications or adaptations are made.
Genes to Cells（2016） Vol.21 Issue 6 p.661-669
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LID201706014001.pdf 818 KB
Institute of Advanced Medical Sciences