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DOI | 10.1038/s41586-020-2294-9 |
Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform | |
Touat, Mehdi1,2,3; Li, Yvonne Y.2,4; Boynton, Adam N.2,5; Spurr, Liam F.2,4; Iorgulescu, J. Bryan4,6; Bohrson, Craig L.7,8; Cortes-Ciriano, Isidro9; Birzu, Cristina3; Geduldig, Jack E.1; Pelton, Kristine1; Lim-Fat, Mary Jane4,10; Pal, Sangita2,4; Ferrer-Luna, Ruben2,4,11; Ramkissoon, Shakti H.11,12,13; Dubois, Frank2,4; Bellamy, Charlotte1; Currimjee, Naomi4; Bonardi, Juliana1; Qian Kenin5; Ho, Patricia5; Malinowski, Seth1; Taquet, Leon1; Jones, Robert E.1; Shetty, Aniket14; Chow, Kin-Hoe14; Sharaf, Radwa11; Pavlick, Dean11; Albacker, Lee A.11; Younan, Nadia3; Baldini, Capucine15; Verreault, Maite16; Giry, Marine16; Guillerm, Erell17; Ammari, Samy18,19; Beuvon, Frederic20; Mokhtari, Karima21; Alentorn, Agusti3; Dehais, Caroline3; Houillier, Caroline3; Laigle-Donadey, Florence3; Psimaras, Dimitri3; Lee, Eudocia Q.4,10; Nayak, Lakshmi4,10; McFaline-Figueroa, J. Ricardo4,10; Carpentier, Alexandre22; Cornu, Philippe22; Capelle, Laurent22; Mathon, Bertrand22; Barnholtz-Sloan, Jill S.23; Chakravarti, Arnab24; Bi, Wenya Linda25; Chiocca, E. Antonio25; Fehnel, Katie Pricola26; Alexandrescu, Sanda27; Chi, Susan N.5,28; Haas-Kogan, Daphne29; Batchelor, Tracy T.4,10; Frampton, Garrett M.11; Alexander, Brian M.11,29; Huang, Raymond Y.30; Ligon, Azra H.6; Coulet, Florence17; Delattre, Jean-Yves3,31; Hoang-Xuan, Khe3; Meredith, David M.1,6; Santagata, Sandro1,6,32,33; Duval, Alex34; Sanson, Marc3,31; Cherniack, Andrew D.2,4; Wen, Patrick Y.4,10; Reardon, David A.4; Marabelle, Aurelien15; Park, Peter J.7; Idbaih, Ahmed3; Beroukhim, Rameen2,4,10; Bandopadhayay, Pratiti2,5,28; Bielle, Franck21; Ligon, Keith L.1,2,6,14,27 | |
2020-04-01 | |
发表期刊 | NATURE
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ISSN | 0028-0836 |
EISSN | 1476-4687 |
出版年 | 2020 |
文章类型 | Article;Early Access |
语种 | 英语 |
国家 | Switzerland; Germany; Russia |
英文关键词 | Reverse genetics has been an indispensable tool to gain insights into viral pathogenesis and vaccine development. The genomes of large RNA viruses, such as those from coronaviruses, are cumbersome to clone and manipulate inEscherichia coliowing to the size and occasional instability of the genome(1-3). Therefore, an alternative rapid and robust reverse-genetics platform for RNA viruses would benefit the research community. Here we show the full functionality of a yeast-based synthetic genomics platform to genetically reconstruct diverse RNA viruses, including members of theCoronaviridae,FlaviviridaeandPneumoviridaefamilies. Viral subgenomic fragments were generated using viral isolates, cloned viral DNA, clinical samples or synthetic DNA, and these fragments were then reassembled in one step inSaccharomyces cerevisiaeusing transformation-associated recombination cloning to maintain the genome as a yeast artificial chromosome. T7 RNA polymerase was then used to generate infectious RNA to rescue viable virus. Using this platform, we were able to engineer and generate chemically synthesized clones of the virus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)(4), which has caused the recent pandemic of coronavirus disease (COVID-19), in only a week after receipt of the synthetic DNA fragments. The technical advance that we describe here facilitates rapid responses to emerging viruses as it enables the real-time generation and functional characterization of evolving RNA virus variants during an outbreak. A yeast-based synthetic genomics platform is used to reconstruct and characterize large RNA viruses from synthetic DNA fragments this technique will facilitate the rapid analysis of RNA viruses, such as SARS-CoV-2, during an outbreak. |
领域 | 地球科学 ; 气候变化 ; 资源环境 |
收录类别 | SCI-E |
WOS记录号 | WOS:000541050900001 |
WOS关键词 | INFECTIOUS CLONE ; IN-VITRO ; CORONAVIRUS ; RNA ; PNEUMONIA ; SYSTEM |
WOS类目 | Multidisciplinary Sciences |
WOS研究方向 | Science & Technology - Other Topics |
引用统计 | |
文献类型 | 期刊论文 |
条目标识符 | http://119.78.100.173/C666/handle/2XK7JSWQ/281324 |
专题 | 地球科学 资源环境科学 气候变化 |
作者单位 | 1.Harvard Med Sch, Dana Farber Canc Inst, Dept Oncol Pathol, Boston, MA 02115 USA; 2.Broad Inst MIT & Harvard, Cambridge, MA 02142 USA; 3.Sorbonne Univ, Hop Univ La Pitie Salpetriere Charles Foix, AP HP,Serv Neurol 2 Mazarin,CNRS,UMR S 1127, Inst Cerveau & Moelle Epiniere,Inserm,ICM, Paris, France; 4.Harvard Med Sch, Dana Farber Canc Inst, Dept Med Oncol, Boston, MA 02115 USA; 5.Dana Farber Boston Childrens Canc & Blood Disorde, Boston, MA 02215 USA; 6.Harvard Med Sch, Brigham & Womens Hosp, Dept Pathol, Boston, MA 02115 USA; 7.Harvard Med Sch, Dept Biomed Informat, Boston, MA 02115 USA; 8.Harvard Med Sch, Bioinformat & Integrat Genom PhD Program, Boston, MA 02115 USA; 9.European Mol Biol Lab, European Bioinformat Inst, Wellcome Genome Campus, Hinxton, England; 10.Harvard Med Sch, Dept Neurol, Brigham & Womens Hosp, Boston, MA 02115 USA; 11.Fdn Med Inc, Cambridge, MA USA; 12.Wake Forest Comprehens Canc Ctr, Winston Salem, NC USA; 13.Wake Forest Sch Med, Dept Pathol, Winston Salem, NC 27101 USA; 14.Dana Farber Canc Inst, Ctr Patient Derived Models, Boston, MA 02115 USA; 15.Univ Paris Saclay, Drug Dev Dept DITEP, INSERM, U1015,Gustave Roussy, Villejuif, France; 16.Sorbonne Univ, Inst Cerveau & Moelle Epiniere, CNRS, UMR S 1127,Inserm,ICM, Paris, France; 17.Hop Univ La Pitie Salpetriere Charles Foix, Dept Genet, Unite Fonct Oncogenet & Angiogenet Mol, Paris, France; 18.Gustave Roussy, Dept Diagnost Radiol, Villejuif, France; 19.Univ Paris Sud, CNRS, IR4M UMR8081, Orsay, France; 20.Univ Paris 05, Hop Cochin, AP HP, Serv Anat & Cytol Pathol, Paris, France; 21.Sorbonne Univ, Hop Univ La Pitie Salpetriere Charles Foix, AP HP,Inst Cerveau & Moelle Epiniere,ICM, Serv Neuropathol Lab Escourolle,CNRS,UMR S 1127, Paris, France; 22.Sorbonne Univ, Hop Univ La Pitie Salpetriere Charles Foix, AP HP,Inst Cerveau & Moelle Epiniere, Serv Neurochirurg,CNRS,UMR S 1127,Inserm,ICM, Paris, France; 23.Case Western Reserve Univ, Sch Med, Dept Populat & Quantitat Hlth Sci, Cleveland, OH USA; 24.Arthur G James Hospi, Ohio State Comprehens Canc Ctr, Dept Radiat Oncol, Columbus, OH USA; 25.Harvard Med Sch, Dept Neurosurg, Brigham & Womens Hosp, Boston, MA 02115 USA; 26.Harvard Med Sch, Dept Neurosurg, Boston Childrens Hosp, Boston, MA 02115 USA; 27.Harvard Med Sch, Dept Pathol, Boston Childrens Hosp, Boston, MA 02115 USA; 28.Harvard Med Sch, Dept Pediat, Boston, MA 02115 USA; 29.Harvard Med Sch, Brigham & Womens Hosp, Dana Farber Canc Inst, Boston Childrens Hosp,Dept Radiat Oncol, Boston, MA 02115 USA; 30.Harvard Med Sch, Dept Radiol, Brigham & Womens Hosp, Dana Farber Canc Inst, Boston, MA 02115 USA; 31.ICM, Onconeurotek Tumor Bank, Inst Cerveau & Moelle Epiniere, Paris, France; 32.Harvard Med Sch, Ludwig Ctr, Boston, MA 02115 USA; 33.Harvard Med Sch, Lab Syst Pharmacol, Boston, MA 02115 USA; 34.Sorbonne Univ, Equipe Labellisee Ligue Natl Canc, Equipe Instabil Microsatell & Canc, Ctr Rech St Antoine,Inserm,UMR938, Paris, France |
推荐引用方式 GB/T 7714 | Touat, Mehdi,Li, Yvonne Y.,Boynton, Adam N.,et al. Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform[J]. NATURE,2020. |
APA | Touat, Mehdi.,Li, Yvonne Y..,Boynton, Adam N..,Spurr, Liam F..,Iorgulescu, J. Bryan.,...&Ligon, Keith L..(2020).Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform.NATURE. |
MLA | Touat, Mehdi,et al."Rapid reconstruction of SARS-CoV-2 using a synthetic genomics platform".NATURE (2020). |
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