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J Integr Plant Biol. 2018 May;60(5):376-381. doi: 10.1111/jipb.12622. Epub 2018 Feb 8.

Multigene editing via CRISPR/Cas9 guided by a single-sgRNA seed in Arabidopsis.

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Research Centre for Plant RNA Signaling, College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou 310036, China.
Warwick-Hangzhou Joint RNA Signaling Laboratory, School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.
Worcester-Hangzhou Joint Molecular Plant Health Laboratory, Institute of Science and the Environment, University of Worcester, WR2 6AJ, UK.


We report that a solo single-guide RNA (sgRNA) seed is capable of guiding Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR -associated 9 (CRISRP/Cas9) to simultaneously edit multiple genes AtRPL10A, AtRPL10B and AtRPL10C in Arabidopsis. Our results also demonstrate that it is possible to use CRISPR/Cas9 technology to create AtRPL10 triple mutants which otherwise cannot be generated by conventional genetic crossing. Compared to other conventional multiplex CRISPR/Cas systems, a single sgRNA seed has the advantage of reducing off-target gene-editing. Such a gene editing system might be also applicable to modify other homologous genes, or even less-homologous sequences for multiple gene-editing in plants and other organisms.

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