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      Genome editing with the CRISPR‐Cas system: an art, ethics and global regulatory perspective

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          Summary

          Over the last three decades, the development of new genome editing techniques, such as ODM, TALENs, ZFNs and the CRISPR‐Cas system, has led to significant progress in the field of plant and animal breeding. The CRISPR‐Cas system is the most versatile genome editing tool discovered in the history of molecular biology because it can be used to alter diverse genomes (e.g. genomes from both plants and animals) including human genomes with unprecedented ease, accuracy and high efficiency. The recent development and scope of CRISPR‐Cas system have raised new regulatory challenges around the world due to moral, ethical, safety and technical concerns associated with its applications in pre‐clinical and clinical research, biomedicine and agriculture. Here, we review the art, applications and potential risks of CRISPR‐Cas system in genome editing. We also highlight the patent and ethical issues of this technology along with regulatory frameworks established by various nations to regulate CRISPR‐Cas‐modified organisms/products.

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          Most cited references129

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          Repair of double-strand breaks induced by CRISPR–Cas9 leads to large deletions and complex rearrangements

          CRISPR-Cas9 is poised to become the gene editing tool of choice in clinical contexts. Thus far, exploration of Cas9-induced genetic alterations has been limited to the immediate vicinity of the target site and distal off-target sequences, leading to the conclusion that CRISPR-Cas9 was reasonably specific. Here we report significant on-target mutagenesis, such as large deletions and more complex genomic rearrangements at the targeted sites in mouse embryonic stem cells, mouse hematopoietic progenitors and a human differentiated cell line. Using long-read sequencing and long-range PCR genotyping, we show that DNA breaks introduced by single-guide RNA/Cas9 frequently resolved into deletions extending over many kilobases. Furthermore, lesions distal to the cut site and crossover events were identified. The observed genomic damage in mitotically active cells caused by CRISPR-Cas9 editing may have pathogenic consequences.
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            Developing a framework for responsible innovation

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              Functional repair of CFTR by CRISPR/Cas9 in intestinal stem cell organoids of cystic fibrosis patients.

              Single murine and human intestinal stem cells can be expanded in culture over long time periods as genetically and phenotypically stable epithelial organoids. Increased cAMP levels induce rapid swelling of such organoids by opening the cystic fibrosis transmembrane conductor receptor (CFTR). This response is lost in organoids derived from cystic fibrosis (CF) patients. Here we use the CRISPR/Cas9 genome editing system to correct the CFTR locus by homologous recombination in cultured intestinal stem cells of CF patients. The corrected allele is expressed and fully functional as measured in clonally expanded organoids. This study provides proof of concept for gene correction by homologous recombination in primary adult stem cells derived from patients with a single-gene hereditary defect. Copyright © 2013 Elsevier Inc. All rights reserved.
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                Author and article information

                Contributors
                dinfany@mail.hzau.edu.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
                30 April 2020
                August 2020
                : 18
                : 8 ( doiID: 10.1111/pbi.v18.8 )
                : 1651-1669
                Affiliations
                [ 1 ] National Key Laboratory of Crop Genetic Improvement Huazhong Agricultural University Wuhan China
                [ 2 ] College of Public Administration Huazhong Agricultural University Wuhan China
                [ 3 ] Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction Ministry of Education Wuhan China
                [ 4 ] Key Laboratory of Horticultural Plant Biology Ministry of Education College of Horticulture and Forestry Sciences Huazhong Agricultural University Wuhan China
                [ 5 ] Hubei Key Laboratory of Plant Pathology College of Plant Sciences and Technology Huazhong Agricultural University Wuhan China
                Author notes
                [*] [* ] Correspondence (Tel +86 131 251 81582; fax +027 872 80016; email dinfany@ 123456mail.hzau.edu.cn )

                [*]

                These two authors contribute equally to this research.

                Author information
                https://orcid.org/0000-0003-0643-2456
                https://orcid.org/0000-0002-1495-9154
                https://orcid.org/0000-0002-9697-7179
                Article
                PBI13383
                10.1111/pbi.13383
                7336378
                32271968
                d6cc9a4e-d509-4701-a62c-a3676e70f941
                © 2020 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/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 20 September 2019
                : 22 February 2020
                : 19 March 2020
                Page count
                Figures: 4, Tables: 1, Pages: 19, Words: 18373
                Funding
                Funded by: Consulting Project of China Academy of Engineering
                Award ID: 2018‐XY‐63
                Funded by: National Natural Science Foundation of China , open-funder-registry 10.13039/501100001809;
                Award ID: 31872077
                Funded by: National Key Research and Development Program of China
                Award ID: 2017YFD0202000
                Categories
                Review
                Review
                Custom metadata
                2.0
                August 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.5 mode:remove_FC converted:06.07.2020

                Biotechnology
                genome editing,crispr‐cas system,ethics,regulations,risks
                Biotechnology
                genome editing, crispr‐cas system, ethics, regulations, risks

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