CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis
Abstract The Xenopus tropicalis (Western clawed frog) is an important amphibian model for genetics, developmental and regenerative biology, due to its diploid genetic background and short generation time. CRISPR-Cas13 and CRISPR interference (CRISPRi) systems have recently been employed to suppress...
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BMC
2025-04-01
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| Series: | Cell & Bioscience |
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| Online Access: | https://doi.org/10.1186/s13578-025-01397-8 |
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| author | Xiao-Lin Lin Yi-Min Zhou Ke Meng Jia-Yi Yang Han Zhang Jin-Hua Lin Hai-Yan Wu Xiao-Yu Wang Hui Zhao Shan-Shan Feng Kyu-Sang Park Dong-Qing Cai Li Zheng Xu-Feng Qi |
| author_facet | Xiao-Lin Lin Yi-Min Zhou Ke Meng Jia-Yi Yang Han Zhang Jin-Hua Lin Hai-Yan Wu Xiao-Yu Wang Hui Zhao Shan-Shan Feng Kyu-Sang Park Dong-Qing Cai Li Zheng Xu-Feng Qi |
| author_sort | Xiao-Lin Lin |
| collection | DOAJ |
| description | Abstract The Xenopus tropicalis (Western clawed frog) is an important amphibian model for genetics, developmental and regenerative biology, due to its diploid genetic background and short generation time. CRISPR-Cas13 and CRISPR interference (CRISPRi) systems have recently been employed to suppress mRNA expression in many organisms such as yeast, plants, and mammalian cells. However, no systematic study of these two systems has been carried out in Xenopus tropicalis. Here, we show that CRISPRi rather than CRISPR-Cas13 is an effective and suitable approach to suppress specific mRNA transcription in Xenopus tropicalis embryos. We demonstrated that CRISPRi composed of dCas9 and KRAB-MeCP2 (dCas9-KM) can efficiently target exogenous and endogenous transcripts in Xenopus tropicalis embryos. Moreover, our data suggest that the new KRAB domain from ZIM3 protein (ZIM3-KRAB, ZIM3K) alone has a comparable transcript targeting capacity in Xenopus tropicalis embryos to the traditional fusion repressor KRAB-MeCP2 in which the KRAB domain from KOX1 protein. In conclusion, our results demonstrate that CRISPRi rather than CRISPR-Cas13 is an efficient knockdown platform to explore specific gene function in Xenopus tropicalis embryos. |
| format | Article |
| id | doaj-art-245d9ff8ebb846c4b9bf36940f338fb5 |
| institution | DOAJ |
| issn | 2045-3701 |
| language | English |
| publishDate | 2025-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Cell & Bioscience |
| spelling | doaj-art-245d9ff8ebb846c4b9bf36940f338fb52025-08-20T03:15:12ZengBMCCell & Bioscience2045-37012025-04-0115111610.1186/s13578-025-01397-8CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalisXiao-Lin Lin0Yi-Min Zhou1Ke Meng2Jia-Yi Yang3Han Zhang4Jin-Hua Lin5Hai-Yan Wu6Xiao-Yu Wang7Hui Zhao8Shan-Shan Feng9Kyu-Sang Park10Dong-Qing Cai11Li Zheng12Xu-Feng Qi13Key Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityDepartment of Hematology, First Affiliated Hospital, Jinan UniversityDivision of Histology & Embryology, School of Medicine, Jinan UniversitySchool of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong KongKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityDepartment of Physiology, Wonju College of Medicine, Yonsei UniversityKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversitySchool of Environmental Science and Engineering, Guangdong University of TechnologyKey Laboratory of Regenerative Medicine of Ministry of Education, Department of Developmental & Regenerative Biology, Jinan UniversityAbstract The Xenopus tropicalis (Western clawed frog) is an important amphibian model for genetics, developmental and regenerative biology, due to its diploid genetic background and short generation time. CRISPR-Cas13 and CRISPR interference (CRISPRi) systems have recently been employed to suppress mRNA expression in many organisms such as yeast, plants, and mammalian cells. However, no systematic study of these two systems has been carried out in Xenopus tropicalis. Here, we show that CRISPRi rather than CRISPR-Cas13 is an effective and suitable approach to suppress specific mRNA transcription in Xenopus tropicalis embryos. We demonstrated that CRISPRi composed of dCas9 and KRAB-MeCP2 (dCas9-KM) can efficiently target exogenous and endogenous transcripts in Xenopus tropicalis embryos. Moreover, our data suggest that the new KRAB domain from ZIM3 protein (ZIM3-KRAB, ZIM3K) alone has a comparable transcript targeting capacity in Xenopus tropicalis embryos to the traditional fusion repressor KRAB-MeCP2 in which the KRAB domain from KOX1 protein. In conclusion, our results demonstrate that CRISPRi rather than CRISPR-Cas13 is an efficient knockdown platform to explore specific gene function in Xenopus tropicalis embryos.https://doi.org/10.1186/s13578-025-01397-8Cas13CRISPRiKnockdownXenopus tropicalis |
| spellingShingle | Xiao-Lin Lin Yi-Min Zhou Ke Meng Jia-Yi Yang Han Zhang Jin-Hua Lin Hai-Yan Wu Xiao-Yu Wang Hui Zhao Shan-Shan Feng Kyu-Sang Park Dong-Qing Cai Li Zheng Xu-Feng Qi CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis Cell & Bioscience Cas13 CRISPRi Knockdown Xenopus tropicalis |
| title | CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis |
| title_full | CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis |
| title_fullStr | CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis |
| title_full_unstemmed | CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis |
| title_short | CRISPR/Cas-mediated mRNA knockdown in the embryos of Xenopus tropicalis |
| title_sort | crispr cas mediated mrna knockdown in the embryos of xenopus tropicalis |
| topic | Cas13 CRISPRi Knockdown Xenopus tropicalis |
| url | https://doi.org/10.1186/s13578-025-01397-8 |
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