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| CRISPR-dCas13-tracing reveals transcriptional memory and limited mRNA export in developing zebrafish embryos | |
| 2023-12-01 | |
| 发表期刊 | GENOME BIOLOGY;
 |
| ISSN | 1474-7596 |
| EISSN | 1474-760X |
| 卷号 | 24期号:1 |
| DOI | 10.1186/s13059-023-02848-6 |
| 摘要 | Background: Understanding gene transcription and mRNA-protein (mRNP) dynamics in single cells in a multicellular organism has been challenging. The catalytically dead CRISPR-Cas13 (dCas13) system has been used to visualize RNAs in live cells without genetic manipulation. We optimize this system to track developmentally expressed mRNAs in zebrafish embryos and to understand features of endogenous transcription kinetics and mRNP export. Results: We report that zygotic microinjection of purified CRISPR-dCas13-fluorescent proteins and modified guide RNAs allows single- and dual-color tracking of developmentally expressed mRNAs in zebrafish embryos from zygotic genome activation (ZGA) until early segmentation period without genetic manipulation. Using this approach, we uncover non-synchronized de novo transcription between inter-alleles, synchronized post-mitotic re-activation in pairs of alleles, and transcriptional memory as an extrinsic noise that potentially contributes to synchronized post-mitotic re-activation. We also reveal rapid dCas13-engaged mRNP movement in the nucleus with a corralled and diffusive motion, but a wide varying range of rate-limiting mRNP export, which can be shortened by Alyref and Nxf1 overexpression. Conclusions: This optimized dCas13-based toolkit enables robust spatial-temporal tracking of endogenous mRNAs and uncovers features of transcription and mRNP motion, providing a powerful toolkit for endogenous RNA visualization in a multicellular developmental organism. |
| 关键词 | De novo transcription Developmental embryos Modified gRNAs mRNP export Post-mitotic Zygotic microinjection |
| 学科门类 | Ecology, Evolution, Behavior and Systematics ; Genetics ; Cell Biology |
| URL | 查看原文 |
| 收录类别 | SCOPUS |
| 语种 | 英语 |
| 原始文献类型 | Article |
| Scopus 记录号 | 2-s2.0-85146630605 |
| 来源库 | SCOPUS |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/286448 |
| 专题 | 生命科学与技术学院 生命科学与技术学院_特聘教授组_陈玲玲组 |
| 作者单位 | 1.State Key Laboratory of Molecular Biology,Shanghai Key Laboratory of Molecular Andrology,CAS Center for Excellence in Molecular Cell Science,Shanghai Institute of Biochemistry and Cell Biology,University of Chinese Academy of Sciences,Chinese Academy of Sciences,320 Yueyang Road,Shanghai,China; 2.CAS Key Laboratory of Computational Biology,Shanghai Institute of Nutrition and Health,University of Chinese Academy of Sciences,Chinese Academy of Sciences,Shanghai,China; 3.National Laboratory of Biomacromolecules,CAS Center for Excellence in Biomacromolecules,Institute of Biophysics,Chinese Academy of Sciences,Beijing,China; 4.College of Life Sciences,University of Chinese Academy of Sciences,Beijing,China; 5.Center for Molecular Medicine,Children’s Hospital,Fudan University and Shanghai Key Laboratory of Medical Epigenetics,International Laboratory of Medical Epigenetics and Metabolism,Ministry of Science and Technology,Institutes of Biomedical Sciences,Fudan University,Shanghai,China; 6.School of Life Science and Technology,ShanghaiTech University,Shanghai,China; 7.School of Life Science,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,Hangzhou,China |
| 推荐引用方式 GB/T 7714 | Huang, Youkui,Gao, Bao-Qing,Meng, Quan,et al. CRISPR-dCas13-tracing reveals transcriptional memory and limited mRNA export in developing zebrafish embryos[J]. GENOME BIOLOGY;,2023,24(1). |
| APA | Huang, Youkui.,Gao, Bao-Qing.,Meng, Quan.,Yang, Liang-Zhong.,Ma, Xu-Kai.,...&Chen, Ling-Ling.(2023).CRISPR-dCas13-tracing reveals transcriptional memory and limited mRNA export in developing zebrafish embryos.GENOME BIOLOGY;,24(1). |
| MLA | Huang, Youkui,et al."CRISPR-dCas13-tracing reveals transcriptional memory and limited mRNA export in developing zebrafish embryos".GENOME BIOLOGY; 24.1(2023). |
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