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Genes & Diseases 2023年第1期10卷 41-44页

作者： Jialong Qi You Zhou Zhikai Zhu Chuanshan Xu Wenjie Chen Key Laboratory of Molecular Target&Clinical Pharmacology and the State&NMPA Key Laboratory of Respiratory Disease School of Pharmaceutical Sciences&the Fifth Affiliated HospitalGuangzhou Medical UniversityGuangzhouGuangdong 511436China Yunnan Digestive Endoscopy Clinical Medical Center Department of GastroenterologyThe First People's Hospital of Yunnan ProvinceKunming 650032China Beijing Tiantan Hospital Capital Medical UniversityBeijing 100070China State Key Laboratory of Respiratory Disease Guangdong-Hongkong-Macao Joint Laboratory of Respiratory Infectious DiseaseGuangzhouGuangdong 510182China Sydney Vital Translational Cancer Research Centre Westbourne StNSW 2065Australia

The adenosine deaminase acting on RNA(ADAR)protein family was well characterized as RNA editing enzymes converting adenosines to inosines(A-to-l)in dsRNA structures.They share a homologous structure including the dsRNA-binding domain and the deaminase domain.ADAR1 function as the primary editing enzyme for A-to-l mutation because of its higher and ubiquitous expression compared to ADAR2 and ADAR3 in eukaryotes.

关键词： ADAR1 adenocarcinoma editing

Enhancing prime editing efficiency and flexibility with tethered and split pegRNAs

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Protein & Cell 2023年第4期14卷 304-308页

作者： Ying Feng Siyuan Liu Qiqin Mo Pengpeng Liu Xiao Xiao Hanhui Ma School of Biotechnology East China University of Science and TechnologyShanghai 200237China Gene editing Center School of Life Science and TechnologyShanghaiTech UniversityShanghai 200135China Department of Molecular Cell and Cancer BiologyUniversity of Massachusetts Medical SchoolWorcesterMA O01655USA

Dear Editor,Most human genetic diseases arise from mutations such as insertion,deletion,or point mutations(Landrum et al.,2016).CRISPRCas system has been repurposed to correct pathogenic mutations in a variety of genetic diseases(Choi et al.,2022).There are many concerns about using CRISPR-mediated double-stranded DNA breaks(DSBs)for therapeutic purposes,primarily due to off-targeted mutations(Kosicki et al.,2018).Nevertheless,base editing cannot correct deletions,insertions,or some point mutations such as transversion mutations.Prime editing has its advantages of precisely correct point mutations,small insertions,or deletions in animal cells(Anzalone et al.,2019)and plants(Lin et al.,2020).However,prime editing efficiency varies among genomic sites or cell types(Chen et al.,2021;Nelson et al.,2022).

关键词： insertion editing alone

Expressing a human RNA demethylase as an assister improves gene-editing efficiency in plants

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Molecular Plant 2024年第3期17卷 363-366页

作者： Mengyan Bai Wenxin Lin Chunyan Peng Peizhe Song Huaqin Kuang Jieni Lin Jieping Zhang Jiyao Wang Bo Chen Huarong Li Fanjiang Kong Guifang Jia Yuefeng Guan Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design Innovative Center of Molecular Genetics and EvolutionSchool of Life SciencesGuangzhou UniversityGuangzhou 510006China College of Agronomy China Agricultural UniversityBeijing 100193China College of Life Sciences Fujian Agriculture and Forestry UniversityFuzhouFujian 350002China Synthetic and Functional Biomolecules Center Bejing National Laboratory for Molecular SciencesKey Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of EducationCollege of Chemistry and Molecular EngineeringPeking UniversityBeijing 100871China Kingagroot Co. Ltd.QingdaoShandong 266000China Tsinghua University-Peking University Joint Center for Life Sciences Bejing 100871China

Dear Editor,Despite myriad successful applications of gene editing in plant functional genomics research and precision breeding,many challenges persist around the efficiency of gene-editing tools for many plant species.For instance,soybean(Glycine max)is a major crop providing oil and protein to human diets and feedstock,but its gene-editing efficiency remains relatively low(Bai et al.,2019).

关键词： editing breeding

High-efficiency gene editing in Anopheles sinensis using ReMOT control

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Insect Science 2024年第1期31卷 307-312页

作者： Xiao-lin Yang Xia Ling Quan Sun Pin-pin Qiu Kai Xiang Jun-feng Hong Shu-lin He Jie Chen Xin Ding Hai Hu Zheng-bo He Cao Zhou Bin Chen Liang Qiao Chongqing Key Laboratory of Vector Insects Institute of Entomology and Molecular BiologyCollege of Life SciencesChongqing Normal UniversityChongqingChina Xunjian Life Science&Technology Co.Ltd. ChongqingChina State Key Laboratory of Resource Insects Southwest UniversityChongqingChina

Dear editor,Functional genomics studies of mosquito vectors are crucial for understanding their physiological behavior,pathogen transmission,and evolution,and for providing essential molecular targets for genetic control(Severson&Behura,201l;Alphey,2014;Ruzzante et al.,2019;Hong et al.,2022).The main methods involve clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing via embryo microinjection(Criscione et al.,2015;Kistler et al.,2015).

关键词： editor editing injection

Base editing-mediated targeted evolution of ACCase for herbicide-resistant rice mutants

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Journal of Integrative Plant Biology 2022年第11期64卷 2029-2032页

作者： Hongzhi Wang Yuxin He Yingying Wang Zuren Li Jiannan Hao Yijiao Song Mugui Wang Jian-Kang Zhu Shanghai Center for Plant Stress Biology and Center for Excellence in Molecular Plant Sciences Chinese Academy of SciencesShanghai 201602China University of Chinese Academy of Sciences Beijing 100049China Hunan Provincial Key Laboratory for Biology and Control of Weeds Hunan Academy of Agricultural SciencesChangsha 410125China College of Life Sciences Nanjing Agricultural UniversityNanjing 210095China Institute of Advanced Biotechnology and School of Life Sciences Southern University of Science and TechnologyShenzhen 518055China Center for Advanced Bioindustry Technologies Chinese Academy of Agricultural SciencesBeijing 100081China

Base editing technologies enable precise base alterations in a target gene without inducing double-stranded breaks,and thus are powerful for targeted gene evolution in vivo(Zhan et al.,2021).The newly evolved cytosine deaminases such as evoAPOBEC1,evoFERNY and evoCDA1,adenine deaminase TadA8e,and a near-protospacer adjacent motif(PAM)less SpRYCas9 variant,have greatly increased base editing efficiency and expanded the editing window in plants(Zhang et al.,2021;Xu et al.,2021c;Tan et al.,2022).Dual cytosine and adenine editors enable concurrent C-to-T and A-to-G conversions and thus enrich the editing outcome.Although plant dual editors have been developed(Li et al.,2020;Xu et al.,2021a),they are rarely applied in evolving plant genes due to low efficiencies.

关键词： evolution enable editing

Multiplex precision gene editing by a surrogate prime editor in rice

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Molecular Plant 2022年第7期15卷 1077-1080页

作者： Huiyuan Li Ziwei Zhu Shaoya Li Jingying Li Lei Yan Chen Zhang Youzhi Ma Lanqin Xia Institute of Crop Sciences(ICS) Chinese Academy of Agricultural Sciences(CAAS)Be11ing 100081China Hainan Yazhou Bay Seed Laboratory/National Nanfan Research Institute(Sanya) Chinese Academy of Agricultural SciencesSanyaHainan Province 572024China

Dear Editor,Development of a multiplex precision gene editing system is highly desirable for pyramiding beneficial alleles in crop improve-ment.Prime editing is a newly developed genome-editing tool that can precisely enable the installation of all 12 nucleotide sub-stitutions,short insertions,and deletions without exogenous DNA donor repair template and double-strand breaks(Anzalone et al.,2019).Among the prime editing systems,prime editor 3(PE3),which consists of a prime editing guide RNA(pegRNA)and an additional nicking single guide RNA(sgRNA)to nick the non-edited strand,enhances editing efficiency.

关键词： editor editing template

Breaking the biotrophic interfacial complex:How genome editing can lead to rice blast resistance

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Molecular Plant 2023年第8期16卷 1243-1245页

作者： Vincent Were Nicholas J.Talbot The Sainsbury Laboratory University of East AngliaNorwich Research ParkNorwich NR47UHUK

Genome editing is a transformational technology—by precisely altering the coding or regulatory sequence of a specific gene,it is now possible to change specific traits within a wide variety of organisms.As a consequence,plant breeding can be taken to a new level of precision,and many countries are moving rapidly to adopt new legislation to permit genome editing(Greenwood et al.,2023).However,genome editing has so far seldom been used to develop disease-resistant crops.This is because plant immunity often depends on single dominant resistance genes,which encode immune receptors that recognize secreted effectors deployed by plant pathogens(Jones and Dangl,2006).In this context,genetic modification has proven to be a much more powerful technology so far to introduce these genes from distinct varieties,or even different species,to develop disease resistant crop species(Greenwood et al.,2023),especially when multiple disease-resistance loci can be introduced together to provide more durable resistance(Luo et al.,2021).Genome editing can be used in an innovative way to resurrect previously effective resistance genes(Contreras et al.,2023),but it has been less effective at introducing new forms of disease resistance to crops.

关键词： breeding resistance editing

Computer-aided engineering of CRISPR-Cas proteins for enhanced human genome editing

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Science China(Life Sciences) 2023年第4期66卷 883-886页

作者： Yangcan Chen Yanping Hu Shengqiu Luo Xinge Wang Bangwei Mao Yi Chen Jing Xu Zhikun Li Qi Zhou Wei Li State Key Laboratory of Stem Cell and Reproductive Biology Institute of ZoologyChinese Academy of SciencesBeijing 100101China University of Chinese Academy of Sciences Beijing 100049China Institute for Stem Cell and Regenerative Medicine Chinese Academy of SciencesBeijing 100101China Bejing Institute for Stem Cell and Regenerative Medicine Beijing 100101China

Dear editor.The remarkably diversified CRISPR-Cas systems in nature have provided unlimited valuable resources to develop genome editing tools that are revolutionizing the fields of biotechnology and medicine.However,due to their microbial origin,the activity of most naturally occurring CRISPR-Cas nucleases is relatively poor in mammalian cells(Ran et al.,2015).Thus,improving the mammalian genome editing efficiency becomes the priority for harnessing more CRISPRCas systems for widespread applications.Rational protein engineering serves as a powerful approach to enhance the catalytic activity of enzymes.Whereas,this approach largely relies on proteinic three-dimension(3D)structure information as guide for design.The fact is that of the large numbers of the CRISPR-Cas systems discovered in recent years,only a small number of them with the 3D structures were reported.To bypass this limitation,here,we report an efficient and simple strategy for rational engineering of Cas nucleases without the need of 3D structure information and successfully optimized nucleases from Cas9 and Cas12 families.

关键词： rational editor editing

Gene editing:from technologies to applications in research and beyond

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Science China(Life Sciences) 2022年第4期65卷 657-659页

作者： Wensheng Wei Caixia Gao Biomedical Pioneering Innovation Center Beijing Advanced Innovation Center for GenomicsPeking-Tsinghua Center for Life SciencesPeking University Genome Editing Research CenterState Key Laboratory of Protein and Plant Gene ResearchSchool of Life SciencesPeking UniversityBeijing100871China State Key Laboratory of Plant Cell and Chromosome Engineering Center for Genome EditingInstitute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing100101China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing100049China

Cutting-edge gene editing technologies enable broadened genomic alternations and accelerate opportunities to use these tools in biomedicine, agriculture, and animal model development. In this issue, Li et al.(2022) reviews the tools of gene editing and highlights key technological developments and its broad applications in biomedicine, hoping to accelerate new discoveries and therapies in biomedicine.

关键词： development editing

Optimized prime editing efficiently generates glyphosate-resistant rice plants carrying homozygous TAP-IVS mutation in EPSPS

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Molecular Plant 2022年第11期15卷 1646-1649页

作者： Yuanyuan Jiang Yiping Chai Dexin Qiao Junya Wang Cuiping Xin Wei Sun Zhenghong Cao Yu Zhang Yun Zhou Xue-Chen Wang Qi-Jun Chen State Key Laboratory of Plant Physiology and Biochemistry College of Biological SciencesChina Agricultural UniversityBeijing 100193China Collaborative Innovation Center of Crop Stress Biology KaifengHenan Province 475004China Institute of Plant Stress Biology School of Life ScienceHenan UniversityKaifeng 475004China Center for Crop Functional Genomics and Molecular Breeding China Agricultural UniversityBeijing 100193China

Dear Editor,Prime editing is a novel and universal CRISPR-Cas-derived precise genome-editing technology and has great potentials for applications in basic plant research and crop molecular breeding(Anzalone et al.,2019).Although low efficiency has restrained the original prime editors(PEs)from being used as a routine tool for precise genome editing in plants,an iterative update of the PEs is removing this obstacle(Lin et al.,2021;Xu et al.,2022).Recently,the Liu group reported three optimization strategies for improving prime editing efficiency(Chen et al.,2021;Nelson et al.,2022).The first strategy is based on engineered prime editing guide RNAs(epegRNAs),which were generated by incorporating structured RNA motifs to the 3′terminus of pegRNAs.This strategy enhances pegRNA stability and prevents degradation of the 3′extension(Nelson et al.,2022).The second strategy is based on the optimized PE2 protein(PEmax),which harbors a SpCas9 variant with increased nuclease activity,an additional nuclear localization signal(NLS)sequences,and a new linker between nCas9 and reverse transcriptase(Chen et al.,2021).The third strategy is based on inhibition of DNA mismatch repair(MMR)in cells(Chen et al.,2021).In this work,we tested the optimized PEs generated with these three strategies in rice,demonstrating that the optimized PEs greatly improved prime editing efficiency in rice.We named the two optimized PEs ePE3max and ePE5max:the former is comprised of the PEmax protein,an epegRNA with evopreQ1 appended to its 3′end,and a nicking sgRNA;the latter is comprised of the ePE3max system and a dominant negative OsMLH1 variant for inhibiting MMR.Using the two optimized PEs,we efficiently generated homozygous and heterozygous T173I,A174V,and P177S(TAP-IVS)mutation in EPSPS in rice,which lays a solid foundation for rice non-transgenic glyphosate-resistance breeding.

关键词： breeding precise editing