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precise editing of a Target Base in the Rice Senome Using a Modified CRISPR/Cas9 System

Molecular Plant 2017年第3期10卷 523-525页

作者： Yuming Lu Jian-Kang Zhu Shanghai Center for Plant Stress Biology Center for Excellence in Molecular Plant Sciences Chinese Academy of Sciences Shanghai 201602 China Department of Horticulture and Landscape Architecture Purdue University West Lafayette IN 47907 USA

CRISPR/Cas9 （clustered regularly interspaced short palindromic repeats/CRISPR-associated Cas9） has been widely used in genome editing in a variety of organisms, including rice （Cong et al., 2013; Feng et al., 2013）.

关键词： precise editing Target Base Rice Genome Using

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Development of Plant Prime-editing Systems for precise Genome editing

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Plant Communications 2020年第3期1卷 124-131页

作者： Rongfang Xu Juan Li Xiaoshuang Liu Tiaofeng Shan Ruiying Qin Pengcheng Wei Key Laboratory of Rice Genetics&Breeding Institute of Rice ResearchAnhui Academy of Agricultural ScienceHefei 230031China

Prime-editing systems have the capability to perform efficient and precise genome editing in human cells.In this study,we first developed a plant prime editor 2(pPE2)system and test its activity by generating a targeted mutation on an HPT^(-ATG) reporter in rice.Our results showed that the pPE2 system could induce programmable editing at different genome sites.In transgenic T0 plants,pPE2-generated mutants occurred with 0%–31.3%frequency,suggesting that the efficiency of pPE2 varied greatly at different genomic sites and with prime-editing guide RNAs of diverse structures.To optimize editing efficiency,guide RNAs were introduced into the pPE2 system following the PE3 and PE3b strategy in human cells.However,at the genomic sites tested in this study,pPE3 systems generated only comparable or even lower editing frequencies.Furthemore,we developed a surrogate pPE2 system by incorporating the HPT^(-ATG) reporter to enrich the prime-edited cells.The nucleotide editing was easily detected in the resistant calli transformed with the surrogate pPE2 system,presumably due to the enhanced screening efficiency of edited cells.Taken together,our results indicate that plant prime-editing systems we developed could provide versatile and flexible editing in rice genome.

关键词： prime editing CRISPR rice precise editing surrogate system

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