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      Simplified adenine base editors improve adenine base editing efficiency in rice

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          Summary

          Adenine base editors ( ABEs) have been exploited to introduce targeted adenine (A) to guanine (G) base conversions in various plant genomes, including rice, wheat and Arabidopsis. However, the ABEs reported thus far are all quite inefficient at many target sites in rice, which hampers their applications in plant genome engineering and crop breeding. Here, we show that unlike in the mammalian system, a simplified base editor ABE‐P1S (Adenine Base Editor‐Plant version 1 Simplified) containing the ecTadA*7.10‐ nSpCas9 (D10A) fusion has much higher editing efficiency in rice compared to the widely used ABE‐P1 consisting of the ecTadA‐ecTadA*7.10‐ nSpCas9 (D10A) fusion. We found that the protein expression level of ABE‐P1S is higher than that of ABE‐P1 in rice calli and protoplasts, which may explain the higher editing efficiency of ABE‐P1S in different rice varieties. Moreover, we demonstrate that the ecTadA*7.10‐ nCas9 fusion can be used to improve the editing efficiency of other ABEs containing SaCas9 or the engineered Sa KKH‐Cas9 variant. These more efficient ABEs will help advance trait improvements in rice and other crops.

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          Most cited references28

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          CRISPR/Cas Genome Editing and Precision Plant Breeding in Agriculture

          Enhanced agricultural production through innovative breeding technology is urgently needed to increase access to nutritious foods worldwide. Recent advances in CRISPR/Cas genome editing enable efficient targeted modification in most crops, thus promising to accelerate crop improvement. Here, we review advances in CRISPR/Cas9 and its variants and examine their applications in plant genome editing and related manipulations. We highlight base-editing tools that enable targeted nucleotide substitutions and describe the various delivery systems, particularly DNA-free methods, that have linked genome editing with crop breeding. We summarize the applications of genome editing for trait improvement, development of techniques for fine-tuning gene regulation, strategies for breeding virus resistance, and the use of high-throughput mutant libraries. We outline future perspectives for genome editing in plant synthetic biology and domestication, advances in delivery systems, editing specificity, homology-directed repair, and gene drives. Finally, we discuss the challenges and opportunities for precision plant breeding and its bright future in agriculture.
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            CRISPR-Based Technologies for the Manipulation of Eukaryotic Genomes.

            The CRISPR-Cas9 RNA-guided DNA endonuclease has contributed to an explosion of advances in the life sciences that have grown from the ability to edit genomes within living cells. In this Review, we summarize CRISPR-based technologies that enable mammalian genome editing and their various applications. We describe recent developments that extend the generality, DNA specificity, product selectivity, and fundamental capabilities of natural CRISPR systems, and we highlight some of the remarkable advancements in basic research, biotechnology, and therapeutics science that these developments have facilitated.
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              • Record: found
              • Abstract: found
              • Article: not found

              Precise base editing in rice, wheat and maize with a Cas9- cytidine deaminase fusion

              Single DNA base pairs are edited in wheat, rice and maize using a Cas9 nickase fusion protein.
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                Author and article information

                Contributors
                jkzhu@sibs.ac.cn
                Journal
                Plant Biotechnol J
                Plant Biotechnol. J
                10.1111/(ISSN)1467-7652
                PBI
                Plant Biotechnology Journal
                John Wiley and Sons Inc. (Hoboken )
                1467-7644
                1467-7652
                19 September 2019
                March 2020
                : 18
                : 3 ( doiID: 10.1111/pbi.v18.3 )
                : 770-778
                Affiliations
                [ 1 ] Shanghai Center for Plant Stress Biology CAS Center of Excellence in Molecular Plant Sciences Chinese Academy of Sciences Shanghai China
                [ 2 ] University of Chinese Academy of Sciences Beijing China
                [ 3 ] College of Life Sciences Nanjing Agricultural University Nanjing China
                [ 4 ] Department of Horticulture and Landscape Architecture Purdue University West Lafayette IN USA
                Author notes
                [*] [* ] Correspondence (Tel (86) 21‐57078201; fax (86) 21‐54920717; email jkzhu@ 123456sibs.ac.cn )
                Author information
                https://orcid.org/0000-0001-5134-731X
                Article
                PBI13244
                10.1111/pbi.13244
                7004905
                31469505
                3f4f7839-eac4-445f-b426-ad772d55b26e
                © 2019 The Authors. Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

                History
                : 30 June 2019
                : 15 August 2019
                : 23 August 2019
                Page count
                Figures: 3, Tables: 2, Pages: 9, Words: 7332
                Funding
                Funded by: Chinese Academy of Sciences , open-funder-registry 10.13039/501100002367;
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                March 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.7.5 mode:remove_FC converted:06.02.2020

                Biotechnology
                abe‐p1s,editing efficiency,ectada deaminase,cas9 variants,rice
                Biotechnology
                abe‐p1s, editing efficiency, ectada deaminase, cas9 variants, rice

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