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
Source PublicationMOLECULAR CELL
ISSN1097-2765
Volume76Issue:6Pages:981-+
Status已发表
DOI10.1016/j.molcel.2019.10.024
AbstractVisualizing 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.
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Indexed BySCI
Language英语
Funding ProjectHoward Hughes Medical Institute[55008728]
WOS Research AreaBiochemistry & Molecular Biology ; Cell Biology
WOS SubjectBiochemistry & Molecular Biology ; Cell Biology
WOS IDWOS:000503436600011
PublisherCELL PRESS
EISSN1097-4164
WOS KeywordNUCLEAR-BODY ; STRESS-RESPONSE ; MESSENGER-RNAS ; GENOMIC LOCI ; TRANSCRIPTION ; PROTEIN ; VISUALIZATION ; PARASPECKLES ; EXPRESSION ; COMPONENTS
Original Document TypeArticle
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Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://kms.shanghaitech.edu.cn/handle/2MSLDSTB/104970
Collection生命科学与技术学院_特聘教授组_陈玲玲组
Corresponding AuthorChen, Ling-Ling
Affiliation1.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
Corresponding Author AffilicationSchool of Life Science and Technology
Recommended Citation
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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