Novel ROSA26 Cre-reporter Knock-in C57BL/6N Mice Exhibiting Green Emission
before and Red Emission after Cre-mediated Recombination |
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Authors: | Yoshikazu Hasegawa Yoko Daitoku Keito Sekiguchi Yoko Tanimoto Saori Mizuno-Iijima Seiya Mizuno Noriko Kajiwara Masatsugu Ema Yoshihiro Miwa Kazuyuki Mekada Atsushi Yoshiki Satoru Takahashi Fumihiro Sugiyama Ken-ichi Yagami |
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Institution: | 1)Laborarory Animal Resource Center, University of Tsukuba, 1–1–1 Tennodai, Tsukuba 305-8575, Japan;2)RIKEN BioResource Center, 3–1–1 Koyadai, Tsukuba 305-0074, Japan |
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Abstract: | The Cre/loxP system is a strategy for controlling temporal and/or spatial gene expression
through genome alteration in mice. As successful Cre/loxP genome alteration depends on
Cre-driver mice, Cre-reporter mice are essential for validation of Cre gene expression
in vivo. In most Cre-reporter mouse strains, although the presence of
reporter product indicates the expression of Cre recombinase, it has remained unclear
whether a lack of reporter signal indicates either no Cre recombinase expression or
insufficient reporter gene promoter activity. We produced a novel ROSA26 knock-in
Cre-reporter C57BL/6N strain exhibiting green emission before and red after Cre-mediated
recombination, designated as strain R26GRR. Ubiquitous green fluorescence and no red
fluorescence were observed in R26GRR mice. To investigate the activation of tdsRed,
EGFP-excised R26GRR, R26RR, mice were produced through the crossing of
C57BL/6N mice with R26GRR/Ayu1-Cre F1 mice. R26RR mice showed extraordinarily
strong red fluorescence in almost all tissues examined, suggesting ubiquitous activation
of the second reporter in all tissues after Cre/loxP recombination. Moreover, endothelial
cell lineage and pancreatic islet-specific expression of red fluorescence were detected in
R26GRR/Tie2-Cre F1 mice and R26GRR /Ins1-Cre F1 mice, respectively.
These results indicated that R26GRR mice are a useful novel Cre-reporter mouse strain. In
addition, R26GRR mice with a pure C57BL/6N background represent a valuable source of
green-to-red photoconvertible cells following Cre/loxP recombination for application in
transplantation studies. The R26GRR mouse strain will be available from RIKEN BioResource
Center (http://www.brc.riken.jp/lab/animal/en/). |
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Keywords: | CAG promoter Cre-reporter mouse EGFP Rosa26 tdsRed |
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