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molecular plant 2013年第3期6卷 729-742页

作者： Qiong Luo Ding Tang mo wang Weixiong Luo Lei Zhang Baoxiang Qin yi shen kejian wang Yafei Li zhukuan cheng College of plant Protection Yunnan Agricultural University Kunming 650201 China state key laboratory of plant genomics and Center for plant Gene Research Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China

MSH5, a meiosis-specific member of the MutS-homolog family, is required for normal level of recombination in budding yeast, mice, Caenorhabditis elegans, and Arabidopsis. Here, we report the identification and characterization of its rice homolog, OsMSH5, and demonstrate its function in rice meiosis. Five independent Osmsh5 mutants exhibited normal vegetative growth and severe sterility. The synaptonemal complex is well installed in Osmsh5, while the chiasma frequency is greatly reduced to approximately 10% of that observed in the wild-type, leading to the homologous non- disjunction and complete sterile phenotype. OsMSH5 is predominantly expressed in panicles. Immunofluorescence studies indicate that OsMSH5 chromosomal localization is limited to the early meiotic prophase I. OsMSH5 can be loaded onto meiotic chromosomes in Oszip4, Osmer3, and hellO. However, those ZMM proteins cannot be localized normally in the absence of OsMSH5. Furthermore, the residual chiasmata were shown to be the least frequent among the zmm mutants, including Osmer3, Oszip4, hellO, and Osmsh5. Taken together, we propose that OsMSH5 functions upstream of OsZIP4, OsMER3, and HEIl0 in class I crossover formation.

关键词： OsMSH5 crossover meiosis rice.

Improving the efficiency of the CRISPR-Cas12a system with tRNA-crRNA arrays

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The Crop Journal 2020年第3期8卷 403-407页

作者： Xixun Hu Xiangbing Meng Jiayang Li kejian wang Hong Yu state key laboratory of Rice biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006ZhejiangChina state key laboratory of plant genomics and National Center for plant Gene Research(beijing) Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China University of chinese academy of sciences Beijing 100049China

CRISPR-Cas12a offers a convenient tool for multiplex genome editing in rice. However, the CRISPR-Cas12a system displays variable editing efficiency across genomic loci, with marked influence by CRISPR RNAs(crRNAs). To improve the efficiency of the CRISPR-Cas12a system for multiplex genome editing, we identified various architectures and expression strategies for crRNAs. Transformation of binary vectors loaded with engineered CRISPR-Cas12a systems into rice calli and subsequent sequencing revealed that a modified tRNA-crRNA array not only efficiently achieved rice multiplex genome editing, but also successfully edited target sites that were not edited by the crRNA array. This improvement contributes to the application of the CRISPR-Cas12a system in plant genome editing, especially for genomic loci that have hitherto been difficult to edit.

关键词： crRNA CRISPR-Cas12a tRNA-crRNA array Genome editing Editing efficiency

ScCas9 recognizes NNG protospacer adjacent motif in genome editing of rice

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Science china(Life sciences) 2020年第3期63卷 450-452页

作者： yibo Xu Xiangbing Meng Junjie wang Baoxiang Qin kejian wang Jiayang Li Chun wang Hong Yu state key laboratory of Rice biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China state key laboratory of plant genomics and National Center for plant Gene Research(beijing) Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeiing 100101China University of chinese academy of sciences Beijing 100049China Agricultural College Guangxi UniversityNanming 530000China

The CRISPR/Cas(clustered regularly in terspaced short palindromic repeats/CRISPR-associated protein)system has been widely used in genome editing,epigenetic modification,and other applications,because it is a precise,inexpensive,powerful and easy-to?use editing tool(Zhang et al.,2019).However,the scope of genome editing is limited by a short protospacer adjacent motif(PAM)specific sequence flanking the target *** pyogenes Cas9(5^Cas9),the most widely used Cas,strongly recognizes NGG PAM and partially recognizes NAG PAM in rice and is restricted to target the sites with these motifs(Anders et al.,2014;Meng et al.,2018).

关键词： CRISPR/Cas spacer editing

Improving the efficiency of prime editing with epegRNAs and high-temperature treatment in rice

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Science china(Life sciences) 2022年第11期65卷 2328-2331页

作者： Jinpeng Zou Xiangbing Meng Qing Liu Meiqi Shang kejian wang Jiayang Li Hong Yu Chun wang state key laboratory of Rice biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China state key laboratory of plant genomics and National Center for Plant Gene Research(Beijing)Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China University of chinese academy of sciences Beijing 100049China Guangdong laboratory of Lingnan modern Agriculture Guangzhou 510642China Hainan Yazhou Bay Seed laboratory Sanya 572025China

Dear Editor,Prime editing(PE)systems are important genome-editing tools developed for mediating arbitrary small DNA insertions,deletions and all 12 base-to-base conversions using a CRISPR-nCas9 fusion of an engineered... 详细信息

关键词： treatment editing

Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice

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Science china(Life sciences) 2018年第1期61卷 122-125页

作者： Xiangbing Meng Xixun Hu Qing Liu Xiaoguang Song Caixia Gao Jiayang Li kejian wang state key laboratory of plant genomics and National Center for plant Gene Research (beijing) Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China state key laboratory of Rtee Btology China National Rice Research Institute Chinese Academy of Agricultural Sciences Hangzhou 310006 China state key laboratory of plant Cell and Chromosome Engineering and Center for Genome Editing Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China University of chinese academy of sciences Beijing 100049 China

Dear Editor,The CRISPR-Cas9（clustered regularly interspaced short palindromic repeats/Cas9）system has been widely used for a variety of applications,including targeted gene knockout,gene insertion,gene replacement and base *** its wide use,the genome editing using CRISPR-Cas9 is performed almost exclusively at sites containing canonical NGG protospacer adjacent motifs（PAMs）.To overcome the PAM constraint of the CRISPR-Cas9 system,many attempts have been made to develop various Cas9 orthologs and v variants with altered PAM specificities （Kleinstiver et al., 2015; Hu et al., 2016）.

关键词： NAG Figure Robust genome editing of CRISPR-Cas9 at NAG PAMs in rice

Expanding the scope of genome editing with SpG and SpRY variants in rice

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Science china(Life sciences) 2021年第10期64卷 1784-1787页

作者： Jun Ren Xiangbing Meng Fengyue Hu Qing Liu Yuexuan Cao Huiying Li Changjie Yan Jiayang Li kejian wang Hong Yu Chun wang state key laboratory of Rice biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China state key laboratory of plant genomics and National Center for plant Gene Research(beijing) Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing 100101China University of chinese academy of sciences Beijing 100049China key laboratory of plant Functional genomics Ministry of EducationYangzhou UniversityYangzhou 225009China

Dear Editor,The clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9(CRISPR/Cas9)system,since it was excavated,has been rapidly developed and sparked a revolution in the genome editing *** principle,CRISPR/Cas9 system relies on the recognition of specific loci on the genome,which is titled the protospacer adjacent motif(PAM).However,the canonical Streptococcus pyogenes Cas9(SpCas9)nuclease only recognizes NGG or NAG PAMs,rendering an inherent obstacle in amplifying the application of CRISPR/Cas9 technology.

关键词： Cas9 nuclease rendering

FED:a web tool for foreign element detection of genome-edited organism

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Science china(Life sciences) 2021年第1期64卷 167-170页

作者： Qing Liu Xiaozhen Jiao Xiangbing Meng Chun wang Cao Xu Zhixi Tian Chuanxiao Xie Genying Li Jiayang Li Hong Yu kejian wang state key laboratory of Rice biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou310006China state key laboratory of plant genomics and National Center for Plant Gene Research(Beijing)Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing100101China state key laboratory of plant Cell and Chromosome Engineering Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChinese Academy of SciencesBeijing100101China National key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop ScienceChinese Academy of Agricultural SciencesBeijing100081China Crop Research institute Shandong Academy of Agricultural SciencesJinan250100China University of chinese academy of sciences Beijing100049China

Dear Editor,Genome editing,especially the newly developed CRISPR technology,is now widely implemented for diverse medical and agricultural applications(Puchta,2018).However,for genome editing,the DNA cassettes encodin... 详细信息

关键词： Genome element

Protein arginine methyltransferase 3 fine-tunes the assembly/disassembly of pre-ribosomes to repress nucleolar stress by interacting with RPS2B in arabidopsis

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molecular plant 2021年第2期14卷 223-236页

作者： Runlai Hang Zhen wang Chao Yang Lilan Luo Beixin mo Xuemei Chen Jing Sun Chunyan Liu Xiaofeng Cao state key laboratory of plant genomics and National Center for plant Gene Research Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China Guangdong Provincial key laboratory for plant Epigenetics College of Life Sciences and OceanographyShenzhen UniversityShenzhen 518060China Department of Botany and plant sciences Institute of Integrative Genome BiologyUniversity of CaliforniaRiversideRiversideCA 92521USA College of Life sciences University of Chinese Academy of SciencesBeijing 100039China CAS Center for Excellence in molecular plant sciences Chinese Academy of SciencesBeijing 100101China

Ribosome biogenesis,which takes place mainly in the nucleolus,involves coordinated expression of preribosomal RNAs(pre-rRNAs)and ribosomal proteins,pre-rRNA processing,and subunit assembly with the aid of numerous assembly *** previous study showed that the Arabidopsis thaliana protein arginine methyltransferase AtPRMT3 regulates pre-rRNA processing;however,the underlying molecular mechanism remains ***,we report that AtPRMT3 interacts with Ribosomal Protein S2(RPS2),facilitating processing of the 90S/Small Subunit(SSU)processome and repressing nucleolar *** isolated an intragenic suppressor of atprmt3-2,which rescues the developmental defects of atprmt3-2 while produces a putative truncated AtPRMT3 protein bearing the entire N-terminus but lacking an intact enzymatic activity domain We further identified RPS2 as an interacting partner of AtPRMT3,and found that loss-of-function rps2a2b mutants were phenotypically reminiscent of atprmt3,showing pleiotropic developmental defects and aberrant pre-rRNA ***2B binds directly to pre-rRNAs in the nucleus,and such binding is enhanced in ***,multiple components of the 90S/SSU processome were more enriched by RPS2B in atprmt3-2,which accounts for early pre-rRNA processing defects and results in nucleolar ***,our study uncovered a novel mechanism by which AtPRMT3 cooperates with RPS2B to facilitate the dynamic assembly/disassembly of the 90S/SSU processome during ribosome biogenesis and repress nucleolar stress.

关键词： AtPRMT3 RPS2 ribosome assembly pre-rRNA processing 90S/SSU processome nucleolar stress

STEME系统:一种助力体内定向进化的新工具

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遗传 2020年第3期42卷 231-235页

作者：胡风越王克剑中国水稻研究所水稻生物学国家重点实验室杭州310006

通过定向进化(directed evolution)可以快速进行蛋白工程改良及重要基因功能研究,以获得新型农艺性状突变体。近期,中国科学院遗传与发育生物学研究所高彩霞团队和李家洋团队合作构建了新型的饱和靶向内源诱变编辑器(saturated targeted... 详细信息

通过定向进化(directed evolution)可以快速进行蛋白工程改良及重要基因功能研究,以获得新型农艺性状突变体。近期,中国科学院遗传与发育生物学研究所高彩霞团队和李家洋团队合作构建了新型的饱和靶向内源诱变编辑器(saturated targeted endogenous mutagenesis editors, STEMEs),并在植物中实现了基因的定向进化和功能筛选。该系统融合了现有的2种单碱基编辑技术,成功实现在植物体内同时诱导C:G>T:A、A:T>G:C双碱基编辑,通过靶向OsACC羧基转移酶结构域编码序列定向进化出水稻除草剂抗性植株。这种在体内进行基因定向进化的新方法,对于今后农作物重要农艺性状的筛选和功能基因研究具有重要作用。本文对STEME系统的组成、编辑效率和应用原理进行介绍,并与已有的定向进化方法进行比较,为加速作物种质资源创新研究提供参考。

关键词： STEME 双碱基编辑定向进化种质创新