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Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression
The Mucorales fungal genus Rhizopus is used for the industrial production of organic acids, enzymes and fermented foods. The metabolic engineering efficiency of Rhizopus could be improved using gene manipulation; however, exogenous DNA rarely integrates into the host genome. Consequently, a genetic...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8863565/ https://www.ncbi.nlm.nih.gov/pubmed/35137045 http://dx.doi.org/10.1093/femsle/fnac010 |
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author | Tsuboi, Yuichi Sakuma, Tetsushi Yamamoto, Takashi Horiuchi, Hiroyuki Takahashi, Fumikazu Igarashi, Kazuaki Hagihara, Hiroshi Takimura, Yasushi |
author_facet | Tsuboi, Yuichi Sakuma, Tetsushi Yamamoto, Takashi Horiuchi, Hiroyuki Takahashi, Fumikazu Igarashi, Kazuaki Hagihara, Hiroshi Takimura, Yasushi |
author_sort | Tsuboi, Yuichi |
collection | PubMed |
description | The Mucorales fungal genus Rhizopus is used for the industrial production of organic acids, enzymes and fermented foods. The metabolic engineering efficiency of Rhizopus could be improved using gene manipulation; however, exogenous DNA rarely integrates into the host genome. Consequently, a genetic tool for Mucorales fungi needs to be developed. Recently, programmable nucleases that generate DNA double-strand breaks (DSBs) at specific genomic loci have been used for genome editing in various organisms. In this study, we examined gene disruption in Rhizopus oryzae using transcription activator-like effector nucleases (TALENs), with and without exonuclease overexpression. TALENs with an overexpressing exonuclease induced DSBs, followed by target site deletions. Although DSBs are repaired mainly by nonhomologous end joining in most organisms, our results suggested that in R. oryzae microhomology-mediated end joining was the major DSB repair system. Our gene manipulation method using TALENs coupled with exonuclease overexpression contributes to basic scientific knowledge and the metabolic engineering of Rhizopus. |
format | Online Article Text |
id | pubmed-8863565 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-88635652022-02-23 Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression Tsuboi, Yuichi Sakuma, Tetsushi Yamamoto, Takashi Horiuchi, Hiroyuki Takahashi, Fumikazu Igarashi, Kazuaki Hagihara, Hiroshi Takimura, Yasushi FEMS Microbiol Lett Research Letter The Mucorales fungal genus Rhizopus is used for the industrial production of organic acids, enzymes and fermented foods. The metabolic engineering efficiency of Rhizopus could be improved using gene manipulation; however, exogenous DNA rarely integrates into the host genome. Consequently, a genetic tool for Mucorales fungi needs to be developed. Recently, programmable nucleases that generate DNA double-strand breaks (DSBs) at specific genomic loci have been used for genome editing in various organisms. In this study, we examined gene disruption in Rhizopus oryzae using transcription activator-like effector nucleases (TALENs), with and without exonuclease overexpression. TALENs with an overexpressing exonuclease induced DSBs, followed by target site deletions. Although DSBs are repaired mainly by nonhomologous end joining in most organisms, our results suggested that in R. oryzae microhomology-mediated end joining was the major DSB repair system. Our gene manipulation method using TALENs coupled with exonuclease overexpression contributes to basic scientific knowledge and the metabolic engineering of Rhizopus. Oxford University Press 2022-02-08 /pmc/articles/PMC8863565/ /pubmed/35137045 http://dx.doi.org/10.1093/femsle/fnac010 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of FEMS. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Research Letter Tsuboi, Yuichi Sakuma, Tetsushi Yamamoto, Takashi Horiuchi, Hiroyuki Takahashi, Fumikazu Igarashi, Kazuaki Hagihara, Hiroshi Takimura, Yasushi Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression |
title | Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression |
title_full | Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression |
title_fullStr | Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression |
title_full_unstemmed | Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression |
title_short | Gene manipulation in the Mucorales fungus Rhizopus oryzae using TALENs with exonuclease overexpression |
title_sort | gene manipulation in the mucorales fungus rhizopus oryzae using talens with exonuclease overexpression |
topic | Research Letter |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8863565/ https://www.ncbi.nlm.nih.gov/pubmed/35137045 http://dx.doi.org/10.1093/femsle/fnac010 |
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