One-step Cre-loxP organism creation by TAx9
Abstract The creation of organisms with Cre-loxP conditional gene recombination systems often faces challenges, particularly when creating the initial (F0) generation with both a Cre recombinase and a DNA site flanked by loxP elements (floxed site). The primary reason is that it is difficult to synt...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-03-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-07759-9 |
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| author | Martin Miguel Casco-Robles Tomoki Echigoya Takeaki Shimazaki Yuri Murakami Masaya Hirano Fumiaki Maruo Seiya Mizuno Satoru Takahashi Chikafumi Chiba |
| author_facet | Martin Miguel Casco-Robles Tomoki Echigoya Takeaki Shimazaki Yuri Murakami Masaya Hirano Fumiaki Maruo Seiya Mizuno Satoru Takahashi Chikafumi Chiba |
| author_sort | Martin Miguel Casco-Robles |
| collection | DOAJ |
| description | Abstract The creation of organisms with Cre-loxP conditional gene recombination systems often faces challenges, particularly when creating the initial (F0) generation with both a Cre recombinase and a DNA site flanked by loxP elements (floxed site). The primary reason is that it is difficult to synthesize a single plasmid with both the Cre gene and the floxed site, since Cre-mediated recombination spontaneously occurs when the plasmid is amplified in Escherichia coli bacterial cells. Here, we introduce an artificial nucleic acid sequence TATATATATATATATATA, named TAx9, that enables the integration of both the Cre gene and the floxed site into a single plasmid. TAx9 effectively blocks spontaneous Cre-mediated recombination in E. coli cells. Using this system, we created an F0 generation of transgenic newts and CRISPR-Cas9 knock-in mice with tissue-specific Cre recombination triggered by tamoxifen. TAx9 technology will be a powerful strategy for creating organisms capable of conditional genetic modification in the F0 generation, accelerating various life science research by reducing the time and cost for ultimately establishing and maintaining lines of genetically modified organisms. |
| format | Article |
| id | doaj-art-b6931aa2a5234063a35d138ba856b30b |
| institution | DOAJ |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-b6931aa2a5234063a35d138ba856b30b2025-08-20T02:59:57ZengNature PortfolioCommunications Biology2399-36422025-03-018111010.1038/s42003-025-07759-9One-step Cre-loxP organism creation by TAx9Martin Miguel Casco-Robles0Tomoki Echigoya1Takeaki Shimazaki2Yuri Murakami3Masaya Hirano4Fumiaki Maruo5Seiya Mizuno6Satoru Takahashi7Chikafumi Chiba8Institute of Life and Environmental Sciences, University of TsukubaGraduate School of Science and Technology, University of TsukubaGraduate School of Life and Environmental Sciences, University of TsukubaGraduate School of Science and Technology, University of TsukubaGraduate School of Science and Technology, University of TsukubaInstitute of Life and Environmental Sciences, University of TsukubaLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine, University of TsukubaLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine, University of TsukubaInstitute of Life and Environmental Sciences, University of TsukubaAbstract The creation of organisms with Cre-loxP conditional gene recombination systems often faces challenges, particularly when creating the initial (F0) generation with both a Cre recombinase and a DNA site flanked by loxP elements (floxed site). The primary reason is that it is difficult to synthesize a single plasmid with both the Cre gene and the floxed site, since Cre-mediated recombination spontaneously occurs when the plasmid is amplified in Escherichia coli bacterial cells. Here, we introduce an artificial nucleic acid sequence TATATATATATATATATA, named TAx9, that enables the integration of both the Cre gene and the floxed site into a single plasmid. TAx9 effectively blocks spontaneous Cre-mediated recombination in E. coli cells. Using this system, we created an F0 generation of transgenic newts and CRISPR-Cas9 knock-in mice with tissue-specific Cre recombination triggered by tamoxifen. TAx9 technology will be a powerful strategy for creating organisms capable of conditional genetic modification in the F0 generation, accelerating various life science research by reducing the time and cost for ultimately establishing and maintaining lines of genetically modified organisms.https://doi.org/10.1038/s42003-025-07759-9 |
| spellingShingle | Martin Miguel Casco-Robles Tomoki Echigoya Takeaki Shimazaki Yuri Murakami Masaya Hirano Fumiaki Maruo Seiya Mizuno Satoru Takahashi Chikafumi Chiba One-step Cre-loxP organism creation by TAx9 Communications Biology |
| title | One-step Cre-loxP organism creation by TAx9 |
| title_full | One-step Cre-loxP organism creation by TAx9 |
| title_fullStr | One-step Cre-loxP organism creation by TAx9 |
| title_full_unstemmed | One-step Cre-loxP organism creation by TAx9 |
| title_short | One-step Cre-loxP organism creation by TAx9 |
| title_sort | one step cre loxp organism creation by tax9 |
| url | https://doi.org/10.1038/s42003-025-07759-9 |
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