Improved bacterial recombineering by parallelized protein discovery.

Bacterial Proteins / genetics Escherichia coli / genetics Genetic Engineering Genome, Bacterial Mutation Pseudomonas aeruginosa / genetics Recombination, Genetic
MAGE RecT genome engineering recombineering synthetic biology

Journal

Proceedings of the National Academy of Sciences of the United States of America
ISSN: 1091-6490
Titre abrégé: Proc Natl Acad Sci U S A
Pays: United States
ID NLM: 7505876

Informations de publication

Date de publication:
16 06 2020
Historique:
pubmed: 30 5 2020
medline: 15 9 2020
entrez: 30 5 2020
Statut: ppublish

Résumé

Exploiting bacteriophage-derived homologous recombination processes has enabled precise, multiplex editing of microbial genomes and the construction of billions of customized genetic variants in a single day. The techniques that enable this, multiplex automated genome engineering (MAGE) and directed evolution with random genomic mutations (DIvERGE), are however, currently limited to a handful of microorganisms for which single-stranded DNA-annealing proteins (SSAPs) that promote efficient recombineering have been identified. Thus, to enable genome-scale engineering in new hosts, efficient SSAPs must first be found. Here we introduce a high-throughput method for SSAP discovery that we call "serial enrichment for efficient recombineering" (SEER). By performing SEER in

Identifiants

DOI: 10.1073/pnas.2001588117 PMID: 32467157 PMC: PMC7306799
pubmed: 32467157
pii: 2001588117
doi: 10.1073/pnas.2001588117
pmc: PMC7306799
doi:

Substances chimiques

Bacterial Proteins 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

Langues

eng

Sous-ensembles de citation

IM

Pagination

13689-13698

Déclaration de conflit d'intérêts

Competing interest statement: G.M.C. has related financial interests in EnEvolv, GRO Biosciences, and 64-x. G.M.C., C.J.G., M.J.L., and X.R. have submitted a patent application relating to pieces of this work (WO2017184227A2). T.M.W., G.T.F., and G.M.C. have submitted a patent application related to the improved single-stranded DNA-annealing proteins variants referenced here. A.N. and C.P. have submitted a patent application related to directed evolution with random genomic mutations (DIvERGE) (PCT/EP2017/082574 [WO2018108987] Mutagenizing Intracellular Nucleic Acids).

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Auteurs

Timothy M Wannier (TM)

Department of Genetics, Harvard Medical School, Boston, MA 02115; timothy_wannier@hms.harvard.edu gchurch@genetics.med.harvard.edu.
ORCID: 0000-0002-4165-4751

Akos Nyerges (A)

Department of Genetics, Harvard Medical School, Boston, MA 02115.
Synthetic and Systems Biology Unit, Institute of Biochemistry, Biological Research Centre, Szeged HU-6726, Hungary.
ORCID: 0000-0002-1581-490X

Helene M Kuchwara (HM)

Department of Genetics, Harvard Medical School, Boston, MA 02115.

Márton Czikkely (M)

Synthetic and Systems Biology Unit, Institute of Biochemistry, Biological Research Centre, Szeged HU-6726, Hungary.

Dávid Balogh (D)

Synthetic and Systems Biology Unit, Institute of Biochemistry, Biological Research Centre, Szeged HU-6726, Hungary.

Gabriel T Filsinger (GT)

Department of Genetics, Harvard Medical School, Boston, MA 02115.
ORCID: 0000-0001-6695-2934

Nathaniel C Borders (NC)

Department of Genetics, Harvard Medical School, Boston, MA 02115.

Christopher J Gregg (CJ)

Department of Genetics, Harvard Medical School, Boston, MA 02115.

Marc J Lajoie (MJ)

Department of Genetics, Harvard Medical School, Boston, MA 02115.
ORCID: 0000-0002-0477-5511

Xavier Rios (X)

Department of Genetics, Harvard Medical School, Boston, MA 02115.

Csaba Pál (C)

Synthetic and Systems Biology Unit, Institute of Biochemistry, Biological Research Centre, Szeged HU-6726, Hungary.

George M Church (GM)

Department of Genetics, Harvard Medical School, Boston, MA 02115; timothy_wannier@hms.harvard.edu gchurch@genetics.med.harvard.edu.
ORCID: 0000-0003-3535-2076

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Protéines bactériennes › Improved bacterial recombineering by...
questionsmedicales.fr › Bactéries › Proteobacteria › Gammaproteobacteria › Enterobacteriaceae › Escherichia › Escherichia coli › Improved bacterial recombineering by...
questionsmedicales.fr › Techniques d'investigation › Techniques génétiques › Génie génétique › Improved bacterial recombineering by...
questionsmedicales.fr › Phénomènes génétiques › Structures génétiques › Génome › Génome microbien › Génome bactérien › Improved bacterial recombineering by...
questionsmedicales.fr › Phénomènes génétiques › Variation génétique › Mutation › Improved bacterial recombineering by...
questionsmedicales.fr › Bactéries › Proteobacteria › Gammaproteobacteria › Pseudomonadaceae › Pseudomonas › Pseudomonas aeruginosa › Improved bacterial recombineering by...
questionsmedicales.fr › Phénomènes génétiques › Recombinaison génétique › Improved bacterial recombineering by...