A modular cloning system for standardized assembly of multigene constructs.

The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic element...

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Autores principales:	Ernst Weber, Carola Engler, Ramona Gruetzner, Stefan Werner, Sylvestre Marillonnet
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3f13e5f5be6347998ec5644eed1d05e9
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id	oai:doaj.org-article:3f13e5f5be6347998ec5644eed1d05e9
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spelling	oai:doaj.org-article:3f13e5f5be6347998ec5644eed1d05e92021-11-18T06:58:35ZA modular cloning system for standardized assembly of multigene constructs.1932-620310.1371/journal.pone.0016765https://doaj.org/article/3f13e5f5be6347998ec5644eed1d05e92011-02-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21364738/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic elements in various predefined arrangements. We present here a hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences. This system is based on the ability of type IIS restriction enzymes to assemble multiple DNA fragments in a defined linear order. We constructed a 33 kb DNA molecule containing 11 transcription units made from 44 individual basic modules in only three successive cloning steps. This modular cloning (MoClo) system can be readily automated and will be extremely useful for applications such as gene stacking and metabolic engineering.Ernst WeberCarola EnglerRamona GruetznerStefan WernerSylvestre MarillonnetPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 2, p e16765 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ernst Weber Carola Engler Ramona Gruetzner Stefan Werner Sylvestre Marillonnet A modular cloning system for standardized assembly of multigene constructs.
description	The field of synthetic biology promises to revolutionize biotechnology through the design of organisms with novel phenotypes useful for medicine, agriculture and industry. However, a limiting factor is the ability of current methods to assemble complex DNA molecules encoding multiple genetic elements in various predefined arrangements. We present here a hierarchical modular cloning system that allows the creation at will and with high efficiency of any eukaryotic multigene construct, starting from libraries of defined and validated basic modules containing regulatory and coding sequences. This system is based on the ability of type IIS restriction enzymes to assemble multiple DNA fragments in a defined linear order. We constructed a 33 kb DNA molecule containing 11 transcription units made from 44 individual basic modules in only three successive cloning steps. This modular cloning (MoClo) system can be readily automated and will be extremely useful for applications such as gene stacking and metabolic engineering.
format	article
author	Ernst Weber Carola Engler Ramona Gruetzner Stefan Werner Sylvestre Marillonnet
author_facet	Ernst Weber Carola Engler Ramona Gruetzner Stefan Werner Sylvestre Marillonnet
author_sort	Ernst Weber
title	A modular cloning system for standardized assembly of multigene constructs.
title_short	A modular cloning system for standardized assembly of multigene constructs.
title_full	A modular cloning system for standardized assembly of multigene constructs.
title_fullStr	A modular cloning system for standardized assembly of multigene constructs.
title_full_unstemmed	A modular cloning system for standardized assembly of multigene constructs.
title_sort	modular cloning system for standardized assembly of multigene constructs.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/3f13e5f5be6347998ec5644eed1d05e9
work_keys_str_mv	AT ernstweber amodularcloningsystemforstandardizedassemblyofmultigeneconstructs AT carolaengler amodularcloningsystemforstandardizedassemblyofmultigeneconstructs AT ramonagruetzner amodularcloningsystemforstandardizedassemblyofmultigeneconstructs AT stefanwerner amodularcloningsystemforstandardizedassemblyofmultigeneconstructs AT sylvestremarillonnet amodularcloningsystemforstandardizedassemblyofmultigeneconstructs AT ernstweber modularcloningsystemforstandardizedassemblyofmultigeneconstructs AT carolaengler modularcloningsystemforstandardizedassemblyofmultigeneconstructs AT ramonagruetzner modularcloningsystemforstandardizedassemblyofmultigeneconstructs AT stefanwerner modularcloningsystemforstandardizedassemblyofmultigeneconstructs AT sylvestremarillonnet modularcloningsystemforstandardizedassemblyofmultigeneconstructs
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A modular cloning system for standardized assembly of multigene constructs.

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