CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UFRGS |
Texto Completo: | http://hdl.handle.net/10183/253335 |
Resumo: | Sugarcane is the source of 80% of the sugar and 26% of the bioethanol produced globally. However, its complex, highly polyploid genome (2n = 100 – 120) impedes crop improvement. Here, we report efficient and reproducible gene targeting (GT) in sugarcane, enabling precise co-editing of multiple alleles via template-mediated and homology-directed repair (HDR) of DNA double strand breaks induced by the programmable nuclease CRISPR/Cas9. The evaluation of 146 independently transformed plants from five independent experiments revealed a targeted nucleotide replacement that resulted in both targeted amino acid substitutions W574L and S653I in the acetolactate synthase (ALS) in 11 lines in addition to single, targeted amino acid substitutions W574L or S653I in 25 or 18 lines, respectively. Co-editing of up to three ALS copies/alleles that confer herbicide tolerance was confirmed by Sanger sequencing of cloned long polymerase chain reaction (PCR) amplicons. This work will enable crop improvement by conversion of inferior alleles to superior alleles through targeted nucleotide substitutions. |
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Oz, Mehmet TufanAltpeter, AngelikaKaran, RatnaMerotto Junior, AldoAltpeter, Fredy2023-01-02T05:09:11Z20211664-462xhttp://hdl.handle.net/10183/253335001153389Sugarcane is the source of 80% of the sugar and 26% of the bioethanol produced globally. However, its complex, highly polyploid genome (2n = 100 – 120) impedes crop improvement. Here, we report efficient and reproducible gene targeting (GT) in sugarcane, enabling precise co-editing of multiple alleles via template-mediated and homology-directed repair (HDR) of DNA double strand breaks induced by the programmable nuclease CRISPR/Cas9. The evaluation of 146 independently transformed plants from five independent experiments revealed a targeted nucleotide replacement that resulted in both targeted amino acid substitutions W574L and S653I in the acetolactate synthase (ALS) in 11 lines in addition to single, targeted amino acid substitutions W574L or S653I in 25 or 18 lines, respectively. Co-editing of up to three ALS copies/alleles that confer herbicide tolerance was confirmed by Sanger sequencing of cloned long polymerase chain reaction (PCR) amplicons. This work will enable crop improvement by conversion of inferior alleles to superior alleles through targeted nucleotide substitutions.application/pdfengFrontiers in Genome Editing. Lausanne. Vol. 3 (July 2021), 673566, 12 p.Resistência à pesticidaCana de açúcarResistência genéticaRecombinaçãoRecombinação de homólogosHomologous recombinationCRISPR/Cas9Sugarcane (Saccharum hybrid complex)Genome editingGene targetingHomology directed repairCRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide toleranceEstrangeiroinfo:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFRGSinstname:Universidade Federal do Rio Grande do Sul (UFRGS)instacron:UFRGSTEXT001153389.pdf.txt001153389.pdf.txtExtracted Texttext/plain68046http://www.lume.ufrgs.br/bitstream/10183/253335/2/001153389.pdf.txtda1ca89c9f195b42e011c2a3657126bfMD52ORIGINAL001153389.pdfTexto completo (inglês)application/pdf1601683http://www.lume.ufrgs.br/bitstream/10183/253335/1/001153389.pdf4838798a97af76f09a422824be6d2a2dMD5110183/2533352023-01-04 06:07:26.235591oai:www.lume.ufrgs.br:10183/253335Repositório de PublicaçõesPUBhttps://lume.ufrgs.br/oai/requestopendoar:2023-01-04T08:07:26Repositório Institucional da UFRGS - Universidade Federal do Rio Grande do Sul (UFRGS)false |
dc.title.pt_BR.fl_str_mv |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
title |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
spellingShingle |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance Oz, Mehmet Tufan Resistência à pesticida Cana de açúcar Resistência genética Recombinação Recombinação de homólogos Homologous recombination CRISPR/Cas9 Sugarcane (Saccharum hybrid complex) Genome editing Gene targeting Homology directed repair |
title_short |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
title_full |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
title_fullStr |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
title_full_unstemmed |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
title_sort |
CRISPR/Cas9-mediated multi-allelic gene targeting in sugarcane confers herbicide tolerance |
author |
Oz, Mehmet Tufan |
author_facet |
Oz, Mehmet Tufan Altpeter, Angelika Karan, Ratna Merotto Junior, Aldo Altpeter, Fredy |
author_role |
author |
author2 |
Altpeter, Angelika Karan, Ratna Merotto Junior, Aldo Altpeter, Fredy |
author2_role |
author author author author |
dc.contributor.author.fl_str_mv |
Oz, Mehmet Tufan Altpeter, Angelika Karan, Ratna Merotto Junior, Aldo Altpeter, Fredy |
dc.subject.por.fl_str_mv |
Resistência à pesticida Cana de açúcar Resistência genética Recombinação Recombinação de homólogos |
topic |
Resistência à pesticida Cana de açúcar Resistência genética Recombinação Recombinação de homólogos Homologous recombination CRISPR/Cas9 Sugarcane (Saccharum hybrid complex) Genome editing Gene targeting Homology directed repair |
dc.subject.eng.fl_str_mv |
Homologous recombination CRISPR/Cas9 Sugarcane (Saccharum hybrid complex) Genome editing Gene targeting Homology directed repair |
description |
Sugarcane is the source of 80% of the sugar and 26% of the bioethanol produced globally. However, its complex, highly polyploid genome (2n = 100 – 120) impedes crop improvement. Here, we report efficient and reproducible gene targeting (GT) in sugarcane, enabling precise co-editing of multiple alleles via template-mediated and homology-directed repair (HDR) of DNA double strand breaks induced by the programmable nuclease CRISPR/Cas9. The evaluation of 146 independently transformed plants from five independent experiments revealed a targeted nucleotide replacement that resulted in both targeted amino acid substitutions W574L and S653I in the acetolactate synthase (ALS) in 11 lines in addition to single, targeted amino acid substitutions W574L or S653I in 25 or 18 lines, respectively. Co-editing of up to three ALS copies/alleles that confer herbicide tolerance was confirmed by Sanger sequencing of cloned long polymerase chain reaction (PCR) amplicons. This work will enable crop improvement by conversion of inferior alleles to superior alleles through targeted nucleotide substitutions. |
publishDate |
2021 |
dc.date.issued.fl_str_mv |
2021 |
dc.date.accessioned.fl_str_mv |
2023-01-02T05:09:11Z |
dc.type.driver.fl_str_mv |
Estrangeiro info:eu-repo/semantics/article |
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http://hdl.handle.net/10183/253335 |
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1664-462x |
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001153389 |
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http://hdl.handle.net/10183/253335 |
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eng |
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eng |
dc.relation.ispartof.pt_BR.fl_str_mv |
Frontiers in Genome Editing. Lausanne. Vol. 3 (July 2021), 673566, 12 p. |
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info:eu-repo/semantics/openAccess |
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