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Editing Citrus Genome via SaCas9/sgRNA System
SaCas9/sgRNA, derived from Staphylococcus aureus, is an alternative system for genome editing to Streptococcus pyogenes SpCas9/sgRNA. The smaller SaCas9 recognizes a different protospacer adjacent motif (PAM) sequence from SpCas9. SaCas9/sgRNA has been employed to edit the genomes of Arabidopsis, to...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5732962/ https://www.ncbi.nlm.nih.gov/pubmed/29312390 http://dx.doi.org/10.3389/fpls.2017.02135 |
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author | Jia, Hongge Xu, Jin Orbović, Vladimir Zhang, Yunzeng Wang, Nian |
author_facet | Jia, Hongge Xu, Jin Orbović, Vladimir Zhang, Yunzeng Wang, Nian |
author_sort | Jia, Hongge |
collection | PubMed |
description | SaCas9/sgRNA, derived from Staphylococcus aureus, is an alternative system for genome editing to Streptococcus pyogenes SpCas9/sgRNA. The smaller SaCas9 recognizes a different protospacer adjacent motif (PAM) sequence from SpCas9. SaCas9/sgRNA has been employed to edit the genomes of Arabidopsis, tobacco and rice. In this study, we aimed to test its potential in genome editing of citrus. Transient expression of SaCas9/sgRNA in Duncan grapefruit via Xcc-facilitated agroinfiltration showed it can successfully modify CsPDS and Cs2g12470. Subsequently, binary vector GFP-p1380N-SaCas9/35S-sgRNA1:AtU6-sgRNA2 was developed to edit two target sites of Cs7g03360 in transgenic Carrizo citrange. Twelve GFP-positive Carrizo transformants were successfully established, designated as #Cz1 to #Cz12. Based on targeted next generation sequencing results, the mutation rates for the two targets ranged from 15.55 to 39.13% for sgRNA1 and 49.01 to 79.67% for sgRNA2. Therefore, SaCas9/sgRNA can be used as an alternative tool to SpCas9/sgRNA for citrus genome editing. |
format | Online Article Text |
id | pubmed-5732962 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-57329622018-01-08 Editing Citrus Genome via SaCas9/sgRNA System Jia, Hongge Xu, Jin Orbović, Vladimir Zhang, Yunzeng Wang, Nian Front Plant Sci Plant Science SaCas9/sgRNA, derived from Staphylococcus aureus, is an alternative system for genome editing to Streptococcus pyogenes SpCas9/sgRNA. The smaller SaCas9 recognizes a different protospacer adjacent motif (PAM) sequence from SpCas9. SaCas9/sgRNA has been employed to edit the genomes of Arabidopsis, tobacco and rice. In this study, we aimed to test its potential in genome editing of citrus. Transient expression of SaCas9/sgRNA in Duncan grapefruit via Xcc-facilitated agroinfiltration showed it can successfully modify CsPDS and Cs2g12470. Subsequently, binary vector GFP-p1380N-SaCas9/35S-sgRNA1:AtU6-sgRNA2 was developed to edit two target sites of Cs7g03360 in transgenic Carrizo citrange. Twelve GFP-positive Carrizo transformants were successfully established, designated as #Cz1 to #Cz12. Based on targeted next generation sequencing results, the mutation rates for the two targets ranged from 15.55 to 39.13% for sgRNA1 and 49.01 to 79.67% for sgRNA2. Therefore, SaCas9/sgRNA can be used as an alternative tool to SpCas9/sgRNA for citrus genome editing. Frontiers Media S.A. 2017-12-12 /pmc/articles/PMC5732962/ /pubmed/29312390 http://dx.doi.org/10.3389/fpls.2017.02135 Text en Copyright © 2017 Jia, Xu, Orbović, Zhang and Wang. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Plant Science Jia, Hongge Xu, Jin Orbović, Vladimir Zhang, Yunzeng Wang, Nian Editing Citrus Genome via SaCas9/sgRNA System |
title | Editing Citrus Genome via SaCas9/sgRNA System |
title_full | Editing Citrus Genome via SaCas9/sgRNA System |
title_fullStr | Editing Citrus Genome via SaCas9/sgRNA System |
title_full_unstemmed | Editing Citrus Genome via SaCas9/sgRNA System |
title_short | Editing Citrus Genome via SaCas9/sgRNA System |
title_sort | editing citrus genome via sacas9/sgrna system |
topic | Plant Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5732962/ https://www.ncbi.nlm.nih.gov/pubmed/29312390 http://dx.doi.org/10.3389/fpls.2017.02135 |
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