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A gene network engineering platform for lactic acid bacteria

Recent developments in synthetic biology have positioned lactic acid bacteria (LAB) as a major class of cellular chassis for applications. To achieve the full potential of LAB, one fundamental prerequisite is the capacity for rapid engineering of complex gene networks, such as natural biosynthetic p...

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Detalles Bibliográficos
Autores principales: Kong, Wentao, Kapuganti, Venkata S., Lu, Ting
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770204/
https://www.ncbi.nlm.nih.gov/pubmed/26503255
http://dx.doi.org/10.1093/nar/gkv1093
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author Kong, Wentao
Kapuganti, Venkata S.
Lu, Ting
author_facet Kong, Wentao
Kapuganti, Venkata S.
Lu, Ting
author_sort Kong, Wentao
collection PubMed
description Recent developments in synthetic biology have positioned lactic acid bacteria (LAB) as a major class of cellular chassis for applications. To achieve the full potential of LAB, one fundamental prerequisite is the capacity for rapid engineering of complex gene networks, such as natural biosynthetic pathways and multicomponent synthetic circuits, into which cellular functions are encoded. Here, we present a synthetic biology platform for rapid construction and optimization of large-scale gene networks in LAB. The platform involves a copy-controlled shuttle for hosting target networks and two associated strategies that enable efficient genetic editing and phenotypic validation. By using a nisin biosynthesis pathway and its variants as examples, we demonstrated multiplex, continuous editing of small DNA parts, such as ribosome-binding sites, as well as efficient manipulation of large building blocks such as genes and operons. To showcase the platform, we applied it to expand the phenotypic diversity of the nisin pathway by quickly generating a library of 63 pathway variants. We further demonstrated its utility by altering the regulatory topology of the nisin pathway for constitutive bacteriocin biosynthesis. This work demonstrates the feasibility of rapid and advanced engineering of gene networks in LAB, fostering their applications in biomedicine and other areas.
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spelling pubmed-47702042016-02-29 A gene network engineering platform for lactic acid bacteria Kong, Wentao Kapuganti, Venkata S. Lu, Ting Nucleic Acids Res Methods Online Recent developments in synthetic biology have positioned lactic acid bacteria (LAB) as a major class of cellular chassis for applications. To achieve the full potential of LAB, one fundamental prerequisite is the capacity for rapid engineering of complex gene networks, such as natural biosynthetic pathways and multicomponent synthetic circuits, into which cellular functions are encoded. Here, we present a synthetic biology platform for rapid construction and optimization of large-scale gene networks in LAB. The platform involves a copy-controlled shuttle for hosting target networks and two associated strategies that enable efficient genetic editing and phenotypic validation. By using a nisin biosynthesis pathway and its variants as examples, we demonstrated multiplex, continuous editing of small DNA parts, such as ribosome-binding sites, as well as efficient manipulation of large building blocks such as genes and operons. To showcase the platform, we applied it to expand the phenotypic diversity of the nisin pathway by quickly generating a library of 63 pathway variants. We further demonstrated its utility by altering the regulatory topology of the nisin pathway for constitutive bacteriocin biosynthesis. This work demonstrates the feasibility of rapid and advanced engineering of gene networks in LAB, fostering their applications in biomedicine and other areas. Oxford University Press 2016-02-29 2015-10-25 /pmc/articles/PMC4770204/ /pubmed/26503255 http://dx.doi.org/10.1093/nar/gkv1093 Text en © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Kong, Wentao
Kapuganti, Venkata S.
Lu, Ting
A gene network engineering platform for lactic acid bacteria
title A gene network engineering platform for lactic acid bacteria
title_full A gene network engineering platform for lactic acid bacteria
title_fullStr A gene network engineering platform for lactic acid bacteria
title_full_unstemmed A gene network engineering platform for lactic acid bacteria
title_short A gene network engineering platform for lactic acid bacteria
title_sort gene network engineering platform for lactic acid bacteria
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4770204/
https://www.ncbi.nlm.nih.gov/pubmed/26503255
http://dx.doi.org/10.1093/nar/gkv1093
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