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Registro Completo |
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Biblioteca(s): |
Embrapa Arroz e Feijão. |
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Data corrente: |
02/02/2023 |
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Data da última atualização: |
24/03/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
VANEGAS K. G.; RENDSVIG, J. K. H.; JARCZYSKA, Z. D.; CÔRTES, M. V. de C. B.; VAN ESCH, A. P.; MORERA-GÓMEZ, M.; CONTESINI, F. J.; MORTENSEN, U. H. |
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Afiliação: |
KATHERINA GARCIA VANEGAS, UNIVERSITY OF DENAMARK; JAKOB KRÆMMER HAAR RENDSVIG, UNIVERSITY OF DENMARK; ZOFIA DOROTA JARCZYNSKA, UNIVERSITY OF DENMARK; MARCIO VINICIUS DE C BARROS CORTES, CNPAF; ABEL PETER VAN ESCH, UNIVERSITY OF DENAMARK; MARTÍ MORERA-GÓMEZ, UNIVERSITY OF DENAMARK; FABIANO JARES CONTESINI, UNIVERSITY OF DENAMARK; UFFE HASBRO MORTENSEN, UNIVERSITY OF DENAMARK. |
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Título: |
A Mad7 system for genetic engineering of filamentous fungi. |
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Ano de publicação: |
2023 |
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Fonte/Imprenta: |
Journal of Fungi, v. 9, n. 1, 16, 2023. |
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ISSN: |
2309-608X |
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DOI: |
https://doi.org/10.3390/jof9010016 |
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Idioma: |
Inglês |
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Conteúdo: |
The introduction of CRISPR technologies has revolutionized strain engineering in filamentous fungi. However, its use in commercial applications has been hampered by concerns over intellectual property (IP) ownership, and there is a need for implementing Cas nucleases that are not limited by complex IP constraints. One promising candidate in this context is the Mad7 enzyme, and we here present a versatile Mad7-CRISPR vector-set that can be efficiently used for the genetic engineering of four different Aspergillus species: Aspergillus nidulans, A. niger, A. oryzae and A. campestris, the latter being a species that has never previously been genetically engineered. We successfully used Mad7 to introduce unspecific as well as specific template-directed mutations including gene disruptions, gene insertions and gene deletions. Moreover, we demonstrate that both single-stranded oligonucleotides and PCR fragments equipped with short and long targeting sequences can be used for efficient marker-free gene editing. Importantly, our CRISPR/Mad7 system was functional in both non-homologous end-joining (NHEJ) proficient and deficient strains. Therefore, the newly implemented CRISPR/Mad7 was efficient to promote gene deletions and integrations using different types of DNA repair in four different Aspergillus species, resulting in the expansion of CRISPR toolboxes in fungal cell factories. |
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Palavras-Chave: |
CRISPR; Filamentous fungi; Fungal strain engineering; Mad7. |
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Thesagro: |
Fungo. |
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Thesaurus Nal: |
Aspergillus; Fungi; Genetic engineering. |
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Categoria do assunto: |
X Pesquisa, Tecnologia e Engenharia |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/doc/1151460/1/jof-2023.pdf
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Marc: |
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022 $a2309-608X
024 7 $ahttps://doi.org/10.3390/jof9010016$2DOI
100 1 $aVANEGAS K. G.
245 $aA Mad7 system for genetic engineering of filamentous fungi.$h[electronic resource]
260 $c2023
520 $aThe introduction of CRISPR technologies has revolutionized strain engineering in filamentous fungi. However, its use in commercial applications has been hampered by concerns over intellectual property (IP) ownership, and there is a need for implementing Cas nucleases that are not limited by complex IP constraints. One promising candidate in this context is the Mad7 enzyme, and we here present a versatile Mad7-CRISPR vector-set that can be efficiently used for the genetic engineering of four different Aspergillus species: Aspergillus nidulans, A. niger, A. oryzae and A. campestris, the latter being a species that has never previously been genetically engineered. We successfully used Mad7 to introduce unspecific as well as specific template-directed mutations including gene disruptions, gene insertions and gene deletions. Moreover, we demonstrate that both single-stranded oligonucleotides and PCR fragments equipped with short and long targeting sequences can be used for efficient marker-free gene editing. Importantly, our CRISPR/Mad7 system was functional in both non-homologous end-joining (NHEJ) proficient and deficient strains. Therefore, the newly implemented CRISPR/Mad7 was efficient to promote gene deletions and integrations using different types of DNA repair in four different Aspergillus species, resulting in the expansion of CRISPR toolboxes in fungal cell factories.
650 $aAspergillus
650 $aFungi
650 $aGenetic engineering
650 $aFungo
653 $aCRISPR
653 $aFilamentous fungi
653 $aFungal strain engineering
653 $aMad7
700 1 $aRENDSVIG, J. K. H.
700 1 $aJARCZYSKA, Z. D.
700 1 $aCÔRTES, M. V. de C. B.
700 1 $aVAN ESCH, A. P.
700 1 $aMORERA-GÓMEZ, M.
700 1 $aCONTESINI, F. J.
700 1 $aMORTENSEN, U. H.
773 $tJournal of Fungi$gv. 9, n. 1, 16, 2023.
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Registro original: |
Embrapa Arroz e Feijão (CNPAF) |
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| 1. |  | VANEGAS K. G.; RENDSVIG, J. K. H.; JARCZYSKA, Z. D.; CÔRTES, M. V. de C. B.; VAN ESCH, A. P.; MORERA-GÓMEZ, M.; CONTESINI, F. J.; MORTENSEN, U. H. A Mad7 system for genetic engineering of filamentous fungi. Journal of Fungi, v. 9, n. 1, 16, 2023.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 2 |
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