Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues
Abstract In vivo cell type‐specific genetic recombination based on the Cre‐loxP system has contributed to the understanding of biological processes and diseases. Neuronal nuclei (NeuN)/RBFOX3 is a widely used mature neuron marker in developmental biology and neuroscience. Here, we generated Rbfox3‐i...
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Wiley
2025-04-01
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| Series: | FASEB BioAdvances |
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| Online Access: | https://doi.org/10.1096/fba.2024-00143 |
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| author | Shiho Nishino Misuzu Hashimoto Swapna Paramanya Biswas Natsuki Mikami Yoshikazu Hasegawa Hayate Suzuki Woojin Kang Seiya Mizuno Kazuya Murata |
| author_facet | Shiho Nishino Misuzu Hashimoto Swapna Paramanya Biswas Natsuki Mikami Yoshikazu Hasegawa Hayate Suzuki Woojin Kang Seiya Mizuno Kazuya Murata |
| author_sort | Shiho Nishino |
| collection | DOAJ |
| description | Abstract In vivo cell type‐specific genetic recombination based on the Cre‐loxP system has contributed to the understanding of biological processes and diseases. Neuronal nuclei (NeuN)/RBFOX3 is a widely used mature neuron marker in developmental biology and neuroscience. Here, we generated Rbfox3‐improved Cre (iCre) knock‐in mouse model and investigated the effect of iCre knock‐in into the Rbfox3 gene and Cre recombination activity in the central nervous system (CNS) and peripheral tissues. The knock‐in of internal ribosome entry site (IRES)‐iCre cassette into the Rbfox3 3′ UTR did not affect birth rate, growth, and brain weight. In the adult brain, iCre protein expression was confirmed, whereas RBFOX3 protein expression was partially reduced in the knock‐in mice. Cre recombination analysis using R26GRR fluorescent reporter strain revealed that Rbfox3‐driven iCre‐induced gene recombination in the CNS and heart during embryonic development. In the adult brain, gene recombination was observed in neurons, however, not in other glial cells. In the peripheral tissues, iCre activity was found in the sciatic nerve and in other peripheral tissues, including the heart, bladder, and testis. We validated gene recombination rate in the germline and found that 100% recombination occurred in male germ cells and approximately 50% in female germ cells. Concludingly, Rbfox3‐iCre mice induce genetic recombination in neurons within CNS as well as in some peripheral tissues and germ cells. In addition to establishing a novel Cre mouse line, the findings of this study offer valuable insights into the development and application of mouse tools that utilize the Rbfox3 gene locus. |
| format | Article |
| id | doaj-art-7fa69fc140bc48dfad14c8eb70e19adb |
| institution | OA Journals |
| issn | 2573-9832 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | Wiley |
| record_format | Article |
| series | FASEB BioAdvances |
| spelling | doaj-art-7fa69fc140bc48dfad14c8eb70e19adb2025-08-20T02:09:01ZengWileyFASEB BioAdvances2573-98322025-04-0174n/an/a10.1096/fba.2024-00143Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissuesShiho Nishino0Misuzu Hashimoto1Swapna Paramanya Biswas2Natsuki Mikami3Yoshikazu Hasegawa4Hayate Suzuki5Woojin Kang6Seiya Mizuno7Kazuya Murata8Laboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine University of Tsukuba Tsukuba Ibaraki JapanLaboratory of Biological Chemistry, Faculty of Applied Biological Sciences Gifu University Gifu JapanLaboratory of Biological Chemistry, Faculty of Applied Biological Sciences Gifu University Gifu JapanProgram in Human Biology, School of Integrative and Global Majors University of Tsukuba Tsukuba JapanLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine University of Tsukuba Tsukuba Ibaraki JapanLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine University of Tsukuba Tsukuba Ibaraki JapanLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine University of Tsukuba Tsukuba Ibaraki JapanLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine University of Tsukuba Tsukuba Ibaraki JapanLaboratory Animal Resource Center in Transborder Medical Research Center, Institute of Medicine University of Tsukuba Tsukuba Ibaraki JapanAbstract In vivo cell type‐specific genetic recombination based on the Cre‐loxP system has contributed to the understanding of biological processes and diseases. Neuronal nuclei (NeuN)/RBFOX3 is a widely used mature neuron marker in developmental biology and neuroscience. Here, we generated Rbfox3‐improved Cre (iCre) knock‐in mouse model and investigated the effect of iCre knock‐in into the Rbfox3 gene and Cre recombination activity in the central nervous system (CNS) and peripheral tissues. The knock‐in of internal ribosome entry site (IRES)‐iCre cassette into the Rbfox3 3′ UTR did not affect birth rate, growth, and brain weight. In the adult brain, iCre protein expression was confirmed, whereas RBFOX3 protein expression was partially reduced in the knock‐in mice. Cre recombination analysis using R26GRR fluorescent reporter strain revealed that Rbfox3‐driven iCre‐induced gene recombination in the CNS and heart during embryonic development. In the adult brain, gene recombination was observed in neurons, however, not in other glial cells. In the peripheral tissues, iCre activity was found in the sciatic nerve and in other peripheral tissues, including the heart, bladder, and testis. We validated gene recombination rate in the germline and found that 100% recombination occurred in male germ cells and approximately 50% in female germ cells. Concludingly, Rbfox3‐iCre mice induce genetic recombination in neurons within CNS as well as in some peripheral tissues and germ cells. In addition to establishing a novel Cre mouse line, the findings of this study offer valuable insights into the development and application of mouse tools that utilize the Rbfox3 gene locus.https://doi.org/10.1096/fba.2024-00143bladderCre‐loxPgene recombinationgermlineheartknock‐in mice |
| spellingShingle | Shiho Nishino Misuzu Hashimoto Swapna Paramanya Biswas Natsuki Mikami Yoshikazu Hasegawa Hayate Suzuki Woojin Kang Seiya Mizuno Kazuya Murata Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues FASEB BioAdvances bladder Cre‐loxP gene recombination germline heart knock‐in mice |
| title | Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues |
| title_full | Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues |
| title_fullStr | Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues |
| title_full_unstemmed | Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues |
| title_short | Characterization of the Rbfox3‐IRES‐iCre knock‐in mouse: Revealing gene recombination activity in neural and non‐neural peripheral tissues |
| title_sort | characterization of the rbfox3 ires icre knock in mouse revealing gene recombination activity in neural and non neural peripheral tissues |
| topic | bladder Cre‐loxP gene recombination germline heart knock‐in mice |
| url | https://doi.org/10.1096/fba.2024-00143 |
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