Harnessing stepping-stone hosts to engineer, select, and reboot synthetic bacteriophages in one pot

Cheng, Li1,5,7; Deng, Ziqing1,7,8; Tao, Haoran1,6,7; Song, Wenchen1,7; Xing, Bo1,6,7; Liu, Wenfeng1,7; Kong, Lingxin1,6,7; Yuan, Shengjian2; Ma, Yingfei2; Wu, Yayun3; Huang, Xun3; Peng, Yun4; Wong, Nai-Kei4; Liu, Yingxia4; Wang, Yun1,9,10; Shen, Yue1,9,11; Li, Junhua1,5,7; Xiao, Minfeng1,7

2022-05-23

10.1016/j.crmeth.2022.100217

CELL REPORTS METHODS   影响因子和分区

2667-2375

Advances in synthetic genomics have led to a great demand for genetic manipulation. Trimming any process to simplify and accelerate streamlining of genetic code into life holds great promise for synthesizing and studying organisms. Here, we develop a simple but powerful stepping-stone strategy to promote genome refactoring of viruses in one pot, validated by successful cross-genus and cross-order rebooting of 90 phages infecting 4 orders of popular pathogens. Genomic sequencing suggests that rebooting outcome is associated with gene number and DNA polymerase availability within phage genomes. We integrate recombineering, screening, and rebooting processes in one pot and demonstrate genome assembly and genome editing of phages by stepping-stone hosts in an efficient and economic manner. Under this framework, in vitro assembly, yeast-based assembly, or genetic manipulation of native hosts are not required. As additional stepping-stone hosts are being developed, this framework will open doors for synthetic phages targeting more pathogens and commensals.

ESCI

英语

非南科大

National Key R&D Program of China["SQ2018YFA090004-3","2020YFA0908700"] ; Hunan Health Commission Research Program[20201422] ; Guangdong Provincial Key Laboratory of Genome Read[2017B030301011] ; Shenzhen Engineering Laboratory for Innovative Molecular Diagnostics[DRC-SZ [2016]884] ; Guangdong Provincial Academician Workstation of BGI Synthetic Genomics[2017B090904014]

Biochemistry & Molecular Biology ; Cell Biology

Biochemical Research Methods ; Cell Biology

WOS:000907658200007

ELSEVIER

Web of Science

1.BGI Shenzhen, Shenzhen 518083, Peoples R China
2.Chinese Acad Sci, Shenzhen Inst Synthet Biol, Shenzhen Inst Adv Technol,Guangdong Prov Key Lab, CAS Key Lab Quantitat Engn Biol,Shenzhen Key Lab, Shenzhen 518055, Peoples R China
3.Cent South Univ, Xiangya Hosp, Infect Control Ctr, Changsha 410008, Hunan, Peoples R China
4.Southern Univ Sci & Technol, Shenzhen Key Lab Pathogen & Immun,Shenzhen People, Guangdong Key Lab Diag & Treatment Emerging Infec, State Key Discipline Infect Dis,Hosp 2, Shenzhen 518112, Peoples R China
5.South China Univ Technol, Sch Biol & Biol Engn, Guangzhou 510006, Peoples R China
6.Univ Chinese Acad Sci, Beijing 101408, Peoples R China
7.BGI Shenzhen, Shenzhen Key Lab Unknown Pathogen Identificat, Shenzhen 518083, Peoples R China
8.BGI Beijing, Beijing 100101, Peoples R China
9.BGI Shenzhen, Guangdong Prov Key Lab Genome Read & Write, Shenzhen 518120, Peoples R China
10.BGI Shenzhen, Guangdong Prov Academician Workstn BGI Synthet Ge, Shenzhen 518120, Peoples R China
11.BGI Shenzhen, Shenzhen Engn Lab Innovat Mol Diagnost, Shenzhen 518120, Peoples R China

Cheng, Li,Deng, Ziqing,Tao, Haoran,et al. Harnessing stepping-stone hosts to engineer, select, and reboot synthetic bacteriophages in one pot[J]. CELL REPORTS METHODS,2022,2(5).

Cheng, Li.,Deng, Ziqing.,Tao, Haoran.,Song, Wenchen.,Xing, Bo.,...&Xiao, Minfeng.(2022).Harnessing stepping-stone hosts to engineer, select, and reboot synthetic bacteriophages in one pot.CELL REPORTS METHODS,2(5).

Cheng, Li,et al."Harnessing stepping-stone hosts to engineer, select, and reboot synthetic bacteriophages in one pot".CELL REPORTS METHODS 2.5(2022).