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Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems | |
Yang, Liang-Zhong1; Wang, Yang1; Li, Si-Qi1; Yao, Run-Wen1; Luan, Peng-Fei1; Wu, Huang1; Carmichael, Gordon G.2; Chen, Ling-Ling1,3 | |
2019-12-19 | |
发表期刊 | MOLECULAR CELL |
ISSN | 1097-2765 |
EISSN | 1097-4164 |
卷号 | 76期号:6页码:981-+ |
发表状态 | 已发表 |
DOI | 10.1016/j.molcel.2019.10.024 |
摘要 | Visualizing the location and dynamics of RNAs in live cells is key to understanding their function. Here, we identify two endonuclease-deficient, single-component programmable RNA-guided and RNA-targeting Cas13 RNases (dCasl3s) that allow robust real-time imaging and tracking of RNAs in live cells, even when using single 20- to 27-nt-long guide RNAs. Compared to the aptamer-based MS2-MCP strategy, an optimized dCas13 system is user friendly, does not require genetic manipulation, and achieves comparable RNA-labeling efficiency. We demonstrate that the dCas13 system is capable of labeling NEAT1, SatIII, MUC4, and GCN4 RNAs and allows the study of paraspeckle-associated NEAT1 dynamics. Applying orthogonal dCas13 proteins or combining dCas13 and MS2-MCP allows dual-color imaging of RNAs in single cells. Further combination of dCas13 and dCas9 systems allows simultaneous visualization of genomic DNA and RNA transcripts in living cells. |
URL | 查看原文 |
收录类别 | SCI ; SCIE |
语种 | 英语 |
资助项目 | Howard Hughes Medical Institute[55008728] |
WOS研究方向 | Biochemistry & Molecular Biology ; Cell Biology |
WOS类目 | Biochemistry & Molecular Biology ; Cell Biology |
WOS记录号 | WOS:000503436600011 |
出版者 | CELL PRESS |
WOS关键词 | NUCLEAR-BODY ; STRESS-RESPONSE ; MESSENGER-RNAS ; GENOMIC LOCI ; TRANSCRIPTION ; PROTEIN ; VISUALIZATION ; PARASPECKLES ; EXPRESSION ; COMPONENTS |
原始文献类型 | Article |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | https://kms.shanghaitech.edu.cn/handle/2MSLDSTB/104970 |
专题 | 生命科学与技术学院_特聘教授组_陈玲玲组 |
通讯作者 | Chen, Ling-Ling |
作者单位 | 1.Chinese Acad Sci, CAS Ctr Excellence Mol Cell Sci, Shanghai Inst Biochem & Cell Biol,State Key Lab M, Univ Chinese Acad Sci,Shanghai Key Lab Mol Androl, 320 Yueyang Rd, Shanghai 200031, Peoples R China 2.Univ Connecticut, Hlth Ctr, Stem Cell Inst, Dept Genet & Dev Biol, Farmington, CT 06030 USA 3.ShanghaiTech Univ, Sch Life Sci & Technol, 100 Haike Rd, Shanghai 201210, Peoples R China |
通讯作者单位 | 生命科学与技术学院 |
推荐引用方式 GB/T 7714 | Yang, Liang-Zhong,Wang, Yang,Li, Si-Qi,et al. Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems[J]. MOLECULAR CELL,2019,76(6):981-+. |
APA | Yang, Liang-Zhong.,Wang, Yang.,Li, Si-Qi.,Yao, Run-Wen.,Luan, Peng-Fei.,...&Chen, Ling-Ling.(2019).Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems.MOLECULAR CELL,76(6),981-+. |
MLA | Yang, Liang-Zhong,et al."Dynamic Imaging of RNA in Living Cells by CRISPR-Cas13 Systems".MOLECULAR CELL 76.6(2019):981-+. |
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