Synthetic biology toolbox for nitrogen-fixing soil microbes.
The soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere.
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Format: | Artigo de periódico biblioteca |
Language: | Ingles English |
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2023-12-18
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Subjects: | CRISPR, Cas9, Caixa de ferramentas, Biologia sintética, Edição de genoma, Solo, Nitrogenase, Synthetic biology, Genome, |
Online Access: | http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849 https://doi.org/10.1021/acssynbio.3c00414 |
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dig-alice-doc-11598492023-12-18T13:32:31Z Synthetic biology toolbox for nitrogen-fixing soil microbes. VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. MAYA VENKATARAMAN AUDREY YÑIGEZ-GUTIERREZ VALENTINA INFANTE APRIL MACINTYRE PAULO IVAN FERNANDES JUNIOR, CPATSA JEAN-MICHEL ANÉ BRIAN PFLEGER. CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome The soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere. 2023-12-18T13:32:30Z 2023-12-18T13:32:30Z 2023-12-18 2023 Artigo de periódico ACS Synthetic Biology, v. 12, n. 12, p. 3623-3634, 2023. http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849 https://doi.org/10.1021/acssynbio.3c00414 Ingles en openAccess |
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CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome |
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CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. Synthetic biology toolbox for nitrogen-fixing soil microbes. |
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The soil environment adjacent to plant roots, termed the rhizosphere, is home to a wide variety of microorganisms that can significantly affect the physiology of nearby plants. Microbes in the rhizosphere can provide nutrients, secrete signaling compounds, and inhibit pathogens. These processes could be manipulated with synthetic biology to enhance the agricultural performance of crops grown for food, energy, or environmental remediation, if methods can be implemented in these nonmodel microbes. A common first step for domesticating nonmodel organisms is the development of a set of genetic engineering tools, termed a synthetic biology toolbox. A toolbox comprises transformation protocols, replicating vectors, genome engineering (e.g., CRISPR/Cas9), constitutive and inducible promoter systems, and other gene expression control elements. This work validated synthetic biology toolboxes in three nitrogen-fixing soil bacteria: Azotobacter vinelandii, Stutzerimonas stutzeri (Pseudomonas stutzeri), and a new isolate of Klebsiella variicola. All three organisms were amenable to transformation and reporter protein expression, with several functional inducible systems available for each organism. S. stutzeri and K. variicola showed more reliable plasmid-based expression, resulting in successful Cas9 recombineering to create scarless deletions and insertions. Using these tools, we generated mutants with inducible nitrogenase activity and introduced heterologous genes to produce resorcinol products with relevant biological activity in the rhizosphere. |
author2 |
MAYA VENKATARAMAN |
author_facet |
MAYA VENKATARAMAN VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. |
format |
Artigo de periódico |
topic_facet |
CRISPR Cas9 Caixa de ferramentas Biologia sintética Edição de genoma Solo Nitrogenase Synthetic biology Genome |
author |
VENKATARAMAN, M. YÑIGEZ-GUTIERREZ, A. INFANTE, V. MACINTYRE, A. FERNANDES JUNIOR, P. I. ANÉ, J.-M. PFLEGER, B. |
author_sort |
VENKATARAMAN, M. |
title |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_short |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_full |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_fullStr |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_full_unstemmed |
Synthetic biology toolbox for nitrogen-fixing soil microbes. |
title_sort |
synthetic biology toolbox for nitrogen-fixing soil microbes. |
publishDate |
2023-12-18 |
url |
http://www.alice.cnptia.embrapa.br/alice/handle/doc/1159849 https://doi.org/10.1021/acssynbio.3c00414 |
work_keys_str_mv |
AT venkataramanm syntheticbiologytoolboxfornitrogenfixingsoilmicrobes AT ynigezgutierreza syntheticbiologytoolboxfornitrogenfixingsoilmicrobes AT infantev syntheticbiologytoolboxfornitrogenfixingsoilmicrobes AT macintyrea syntheticbiologytoolboxfornitrogenfixingsoilmicrobes AT fernandesjuniorpi syntheticbiologytoolboxfornitrogenfixingsoilmicrobes AT anejm syntheticbiologytoolboxfornitrogenfixingsoilmicrobes AT pflegerb syntheticbiologytoolboxfornitrogenfixingsoilmicrobes |
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1792482699721572352 |