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A MAD7‐based genome editing system for Escherichia coli
A broad variety of biomolecules is industrially produced in bacteria and yeasts. These microbial expression hosts can be optimized through genetic engineering using CRISPR tools. Here, we designed and characterized such a modular genome editing system based on the Cas12a‐like RNA‐guided nuclease MAD...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
John Wiley and Sons Inc.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10128132/ https://www.ncbi.nlm.nih.gov/pubmed/36929689 http://dx.doi.org/10.1111/1751-7915.14234 |
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author | Mund, Markus Weber, Wadim Degreif, Daniel Schiklenk, Christoph |
author_facet | Mund, Markus Weber, Wadim Degreif, Daniel Schiklenk, Christoph |
author_sort | Mund, Markus |
collection | PubMed |
description | A broad variety of biomolecules is industrially produced in bacteria and yeasts. These microbial expression hosts can be optimized through genetic engineering using CRISPR tools. Here, we designed and characterized such a modular genome editing system based on the Cas12a‐like RNA‐guided nuclease MAD7 in Escherichia coli. This system enables the efficient generation of single nucleotide polymorphisms (SNPs) or gene deletions and can directly be used with donor DNA from benchtop DNA assembly to increase throughput. We combined multiple edits to engineer an E. coli strain with reduced overflow metabolism and increased plasmid yield, highlighting the versatility and industrial applicability of this approach. |
format | Online Article Text |
id | pubmed-10128132 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | John Wiley and Sons Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-101281322023-04-26 A MAD7‐based genome editing system for Escherichia coli Mund, Markus Weber, Wadim Degreif, Daniel Schiklenk, Christoph Microb Biotechnol Regular Issue A broad variety of biomolecules is industrially produced in bacteria and yeasts. These microbial expression hosts can be optimized through genetic engineering using CRISPR tools. Here, we designed and characterized such a modular genome editing system based on the Cas12a‐like RNA‐guided nuclease MAD7 in Escherichia coli. This system enables the efficient generation of single nucleotide polymorphisms (SNPs) or gene deletions and can directly be used with donor DNA from benchtop DNA assembly to increase throughput. We combined multiple edits to engineer an E. coli strain with reduced overflow metabolism and increased plasmid yield, highlighting the versatility and industrial applicability of this approach. John Wiley and Sons Inc. 2023-03-16 /pmc/articles/PMC10128132/ /pubmed/36929689 http://dx.doi.org/10.1111/1751-7915.14234 Text en © 2023 Sanofi Aventis Deutschland GmbH. Microbial Biotechnology published by Applied Microbiology International and John Wiley & Sons Ltd. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ (https://creativecommons.org/licenses/by-nc-nd/4.0/) License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made. |
spellingShingle | Regular Issue Mund, Markus Weber, Wadim Degreif, Daniel Schiklenk, Christoph A MAD7‐based genome editing system for Escherichia coli |
title | A MAD7‐based genome editing system for Escherichia coli
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title_full | A MAD7‐based genome editing system for Escherichia coli
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title_fullStr | A MAD7‐based genome editing system for Escherichia coli
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title_full_unstemmed | A MAD7‐based genome editing system for Escherichia coli
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title_short | A MAD7‐based genome editing system for Escherichia coli
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title_sort | mad7‐based genome editing system for escherichia coli |
topic | Regular Issue |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10128132/ https://www.ncbi.nlm.nih.gov/pubmed/36929689 http://dx.doi.org/10.1111/1751-7915.14234 |
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