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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...

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Autores principales: Jia, Hongge, Xu, Jin, Orbović, Vladimir, Zhang, Yunzeng, Wang, Nian
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2017
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.
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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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