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Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize

CRISPR‐Cas is a powerful DNA double‐strand break technology with wide‐ranging applications in plant genome modification. However, the efficiency of genome editing depends on various factors including plant genetic transformation processes and types of modifications desired. Agrobacterium infection i...

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Autores principales: Peterson, Dave, Barone, Pierluigi, Lenderts, Brian, Schwartz, Chris, Feigenbutz, Lanie, St. Clair, Grace, Jones, Spencer, Svitashev, Sergei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8486252/
https://www.ncbi.nlm.nih.gov/pubmed/33934470
http://dx.doi.org/10.1111/pbi.13613
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author Peterson, Dave
Barone, Pierluigi
Lenderts, Brian
Schwartz, Chris
Feigenbutz, Lanie
St. Clair, Grace
Jones, Spencer
Svitashev, Sergei
author_facet Peterson, Dave
Barone, Pierluigi
Lenderts, Brian
Schwartz, Chris
Feigenbutz, Lanie
St. Clair, Grace
Jones, Spencer
Svitashev, Sergei
author_sort Peterson, Dave
collection PubMed
description CRISPR‐Cas is a powerful DNA double‐strand break technology with wide‐ranging applications in plant genome modification. However, the efficiency of genome editing depends on various factors including plant genetic transformation processes and types of modifications desired. Agrobacterium infection is the preferred method of transformation and delivery of editing components into the plant cell. While this method has been successfully used to generate gene knockouts in multiple crops, precise nucleotide replacement and especially gene insertion into a pre‐defined genomic location remain highly challenging. Here, we report an efficient, selectable marker‐free site‐specific gene insertion in maize using Agrobacterium infection. Advancements in maize transformation and new vector design enabled increase of targeted insertion frequencies by two orders of magnitude in comparison to conventional Agrobacterium‐mediated delivery. Importantly, these advancements allowed not only a significant improvement of the frequency, but also of the quality of generated events. These results further enable the application of genome editing for trait product development in a wide variety of crop species amenable to Agrobacterium‐mediated transformation.
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spelling pubmed-84862522021-10-07 Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize Peterson, Dave Barone, Pierluigi Lenderts, Brian Schwartz, Chris Feigenbutz, Lanie St. Clair, Grace Jones, Spencer Svitashev, Sergei Plant Biotechnol J Research Articles CRISPR‐Cas is a powerful DNA double‐strand break technology with wide‐ranging applications in plant genome modification. However, the efficiency of genome editing depends on various factors including plant genetic transformation processes and types of modifications desired. Agrobacterium infection is the preferred method of transformation and delivery of editing components into the plant cell. While this method has been successfully used to generate gene knockouts in multiple crops, precise nucleotide replacement and especially gene insertion into a pre‐defined genomic location remain highly challenging. Here, we report an efficient, selectable marker‐free site‐specific gene insertion in maize using Agrobacterium infection. Advancements in maize transformation and new vector design enabled increase of targeted insertion frequencies by two orders of magnitude in comparison to conventional Agrobacterium‐mediated delivery. Importantly, these advancements allowed not only a significant improvement of the frequency, but also of the quality of generated events. These results further enable the application of genome editing for trait product development in a wide variety of crop species amenable to Agrobacterium‐mediated transformation. John Wiley and Sons Inc. 2021-07-02 2021-10 /pmc/articles/PMC8486252/ /pubmed/33934470 http://dx.doi.org/10.1111/pbi.13613 Text en © 2021 Corteva Agriscience. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists 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 Research Articles
Peterson, Dave
Barone, Pierluigi
Lenderts, Brian
Schwartz, Chris
Feigenbutz, Lanie
St. Clair, Grace
Jones, Spencer
Svitashev, Sergei
Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
title Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
title_full Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
title_fullStr Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
title_full_unstemmed Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
title_short Advances in Agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
title_sort advances in agrobacterium transformation and vector design result in high‐frequency targeted gene insertion in maize
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8486252/
https://www.ncbi.nlm.nih.gov/pubmed/33934470
http://dx.doi.org/10.1111/pbi.13613
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