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      Use of CRISPR systems in plant genome editing: toward new opportunities in agriculture

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      Emerging Topics in Life Sciences
      Portland Press Ltd.

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          Abstract

          Initially discovered in bacteria and archaea, CRISPR–Cas9 is an adaptive immune system found in prokaryotes. In 2012, scientists found a way to use it as a genome editing tool. In 2013, its application in plants was successfully achieved. This breakthrough has opened up many new opportunities for researchers, including the opportunity to gain a better understanding of plant biological systems more quickly. The present study reviews agricultural applications related to the use of CRISPR systems in plants from 52 peer-reviewed articles published since 2014. Based on this literature review, the main use of CRISPR systems is to achieve improved yield performance, biofortification, biotic and abiotic stress tolerance, with rice (Oryza sativa) being the most studied crop.

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          Most cited references54

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          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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            Efficient and transgene-free genome editing in wheat through transient expression of CRISPR/Cas9 DNA or RNA

            Editing plant genomes is technically challenging in hard-to-transform plants and usually involves transgenic intermediates, which causes regulatory concerns. Here we report two simple and efficient genome-editing methods in which plants are regenerated from callus cells transiently expressing CRISPR/Cas9 introduced as DNA or RNA. This transient expression-based genome-editing system is highly efficient and specific for producing transgene-free and homozygous wheat mutants in the T0 generation. We demonstrate our protocol to edit genes in hexaploid bread wheat and tetraploid durum wheat, and show that we are able to generate mutants with no detectable transgenes. Our methods may be applicable to other plant species, thus offering the potential to accelerate basic and applied plant genome-engineering research.
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              Efficient DNA-free genome editing of bread wheat using CRISPR/Cas9 ribonucleoprotein complexes

              Substantial efforts are being made to optimize the CRISPR/Cas9 system for precision crop breeding. The avoidance of transgene integration and reduction of off-target mutations are the most important targets for optimization. Here, we describe an efficient genome editing method for bread wheat using CRISPR/Cas9 ribonucleoproteins (RNPs). Starting from RNP preparation, the whole protocol takes only seven to nine weeks, with four to five independent mutants produced from 100 immature wheat embryos. Deep sequencing reveals that the chance of off-target mutations in wheat cells is much lower in RNP mediated genome editing than in editing with CRISPR/Cas9 DNA. Consistent with this finding, no off-target mutations are detected in the mutant plants. Because no foreign DNA is used in CRISPR/Cas9 RNP mediated genome editing, the mutants obtained are completely transgene free. This method may be widely applicable for producing genome edited crop plants and has a good prospect of being commercialized.
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                Author and article information

                Journal
                Emerging Topics in Life Sciences
                Emerg. Top. Life Sci.
                Portland Press Ltd.
                2397-8554
                2397-8562
                November 10 2017
                November 10 2017
                November 10 2017
                November 10 2017
                : 1
                : 2
                : 169-182
                Article
                10.1042/ETLS20170085
                0ad799aa-c640-484a-b40e-5664f1a75572
                © 2017

                https://creativecommons.org/licenses/by/4.0/

                History

                Social policy & Welfare,Medicine,Biochemistry,Ecology,Environmental studies,Life sciences

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