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CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato

Genome editing is now widely used in plant science for both basic research and molecular crop breeding. The clustered regularly interspaced short palindromic repeats (CRISPR) technology, through its precision, high efficiency and versatility, allows for editing of many sites in plant genomes. This s...

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Autores principales: Veillet, Florian, Kermarrec, Marie-Paule, Chauvin, Laura, Chauvin, Jean-Eric, Nogué, Fabien
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7430721/
https://www.ncbi.nlm.nih.gov/pubmed/32804931
http://dx.doi.org/10.1371/journal.pone.0235942
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author Veillet, Florian
Kermarrec, Marie-Paule
Chauvin, Laura
Chauvin, Jean-Eric
Nogué, Fabien
author_facet Veillet, Florian
Kermarrec, Marie-Paule
Chauvin, Laura
Chauvin, Jean-Eric
Nogué, Fabien
author_sort Veillet, Florian
collection PubMed
description Genome editing is now widely used in plant science for both basic research and molecular crop breeding. The clustered regularly interspaced short palindromic repeats (CRISPR) technology, through its precision, high efficiency and versatility, allows for editing of many sites in plant genomes. This system has been highly successful to produce knock-out mutants through the introduction of frameshift mutations due to error-prone repair pathways. Nevertheless, recent new CRISPR-based technologies such as base editing and prime editing can generate precise and on demand nucleotide conversion, allowing for fine-tuning of protein function and generating gain-of-function mutants. However, genome editing through CRISPR systems still have some drawbacks and limitations, such as the PAM restriction and the need for more diversity in CRISPR tools to mediate different simultaneous catalytic activities. In this study, we successfully used the CRISPR-Cas9 system from Staphylococcus aureus (SaCas9) for the introduction of frameshift mutations in the tetraploid genome of the cultivated potato (Solanum tuberosum). We also developed a S. aureus-cytosine base editor that mediate nucleotide conversions, allowing for precise modification of specific residues or regulatory elements in potato. Our proof-of-concept in potato expand the plant dicot CRISPR toolbox for biotechnology and precision breeding applications.
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spelling pubmed-74307212020-08-20 CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato Veillet, Florian Kermarrec, Marie-Paule Chauvin, Laura Chauvin, Jean-Eric Nogué, Fabien PLoS One Research Article Genome editing is now widely used in plant science for both basic research and molecular crop breeding. The clustered regularly interspaced short palindromic repeats (CRISPR) technology, through its precision, high efficiency and versatility, allows for editing of many sites in plant genomes. This system has been highly successful to produce knock-out mutants through the introduction of frameshift mutations due to error-prone repair pathways. Nevertheless, recent new CRISPR-based technologies such as base editing and prime editing can generate precise and on demand nucleotide conversion, allowing for fine-tuning of protein function and generating gain-of-function mutants. However, genome editing through CRISPR systems still have some drawbacks and limitations, such as the PAM restriction and the need for more diversity in CRISPR tools to mediate different simultaneous catalytic activities. In this study, we successfully used the CRISPR-Cas9 system from Staphylococcus aureus (SaCas9) for the introduction of frameshift mutations in the tetraploid genome of the cultivated potato (Solanum tuberosum). We also developed a S. aureus-cytosine base editor that mediate nucleotide conversions, allowing for precise modification of specific residues or regulatory elements in potato. Our proof-of-concept in potato expand the plant dicot CRISPR toolbox for biotechnology and precision breeding applications. Public Library of Science 2020-08-17 /pmc/articles/PMC7430721/ /pubmed/32804931 http://dx.doi.org/10.1371/journal.pone.0235942 Text en © 2020 Veillet et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Veillet, Florian
Kermarrec, Marie-Paule
Chauvin, Laura
Chauvin, Jean-Eric
Nogué, Fabien
CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato
title CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato
title_full CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato
title_fullStr CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato
title_full_unstemmed CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato
title_short CRISPR-induced indels and base editing using the Staphylococcus aureus Cas9 in potato
title_sort crispr-induced indels and base editing using the staphylococcus aureus cas9 in potato
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7430721/
https://www.ncbi.nlm.nih.gov/pubmed/32804931
http://dx.doi.org/10.1371/journal.pone.0235942
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