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Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato
CRISPR/Cas9-mediated genome editing technology has been widely used for the study of gene function in crops, but the differences between species have led to widely varying genome editing efficiencies. The present study utilized a potato hairy root genetic transformation system and incorporated a rap...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
PeerJ Inc.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10399558/ https://www.ncbi.nlm.nih.gov/pubmed/37547711 http://dx.doi.org/10.7717/peerj.15771 |
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author | Ye, Mingwang Yao, Mengfan Li, Canhui Gong, Ming |
author_facet | Ye, Mingwang Yao, Mengfan Li, Canhui Gong, Ming |
author_sort | Ye, Mingwang |
collection | PubMed |
description | CRISPR/Cas9-mediated genome editing technology has been widely used for the study of gene function in crops, but the differences between species have led to widely varying genome editing efficiencies. The present study utilized a potato hairy root genetic transformation system and incorporated a rapid assay with GFP as a screening marker. The results clearly demonstrated that salt and osmotic stress induced by NaCl (10 to 50 mM) and mannitol (50 to 200 mM) treatments significantly increased the positive rates of genetic transformation mediated by A. rhizogenes and the editing efficiency of the CRISPR/Cas9-mediated genome editing system in potato. However, it was observed that the regeneration of potato roots was partially inhibited as a result. The analysis of CRISPR/Cas9-mediated mutation types revealed that chimeras accounted for the largest proportion, ranging from 62.50% to 100%. Moreover, the application of salt and osmotic stress resulted in an increased probability of null mutations in potato. Notably, the highest rate of null mutations, reaching 37.5%, was observed at a NaCl concentration of 10 mM. Three potential off-target sites were sequenced and no off-targeting was found. In conclusion, the application of appropriate salt and osmotic stress significantly improved the editing efficiency of the CRISPR/Cas9-mediated genome editing system in potato, with no observed off-target effects. However, there was a trade-off as the regeneration of potato roots was partially inhibited. Overall, these findings present a new and convenient approach to enhance the genome editing efficiency of the CRISPR/Cas9-mediated gene editing system in potato. |
format | Online Article Text |
id | pubmed-10399558 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | PeerJ Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-103995582023-08-04 Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato Ye, Mingwang Yao, Mengfan Li, Canhui Gong, Ming PeerJ Agricultural Science CRISPR/Cas9-mediated genome editing technology has been widely used for the study of gene function in crops, but the differences between species have led to widely varying genome editing efficiencies. The present study utilized a potato hairy root genetic transformation system and incorporated a rapid assay with GFP as a screening marker. The results clearly demonstrated that salt and osmotic stress induced by NaCl (10 to 50 mM) and mannitol (50 to 200 mM) treatments significantly increased the positive rates of genetic transformation mediated by A. rhizogenes and the editing efficiency of the CRISPR/Cas9-mediated genome editing system in potato. However, it was observed that the regeneration of potato roots was partially inhibited as a result. The analysis of CRISPR/Cas9-mediated mutation types revealed that chimeras accounted for the largest proportion, ranging from 62.50% to 100%. Moreover, the application of salt and osmotic stress resulted in an increased probability of null mutations in potato. Notably, the highest rate of null mutations, reaching 37.5%, was observed at a NaCl concentration of 10 mM. Three potential off-target sites were sequenced and no off-targeting was found. In conclusion, the application of appropriate salt and osmotic stress significantly improved the editing efficiency of the CRISPR/Cas9-mediated genome editing system in potato, with no observed off-target effects. However, there was a trade-off as the regeneration of potato roots was partially inhibited. Overall, these findings present a new and convenient approach to enhance the genome editing efficiency of the CRISPR/Cas9-mediated gene editing system in potato. PeerJ Inc. 2023-07-31 /pmc/articles/PMC10399558/ /pubmed/37547711 http://dx.doi.org/10.7717/peerj.15771 Text en © 2023 Ye et al. https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited. |
spellingShingle | Agricultural Science Ye, Mingwang Yao, Mengfan Li, Canhui Gong, Ming Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato |
title | Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato |
title_full | Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato |
title_fullStr | Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato |
title_full_unstemmed | Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato |
title_short | Salt and osmotic stress can improve the editing efficiency of CRISPR/Cas9-mediated genome editing system in potato |
title_sort | salt and osmotic stress can improve the editing efficiency of crispr/cas9-mediated genome editing system in potato |
topic | Agricultural Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10399558/ https://www.ncbi.nlm.nih.gov/pubmed/37547711 http://dx.doi.org/10.7717/peerj.15771 |
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