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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

Regulation of Rice Tillering by RNA-Directed DNA Methylation at Miniature Inverted-Repeat Transposable Elements

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Molecular Plant 2020年第6期13卷 851-863页

作者： Le Xu Kun Yuan meng Yuan xiangbing meng Min Chen Jianguo Wu Jiayang Li Yijun Qi Center for Plant Biology School of Life SciencesTsinghua UniversityBeijing 100084China Tsinghua-Peking Center for Life Sciences Beijing 100084China Key Laboratory of Plant Genomics and National Center for Plant Gene Research Institute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China Vector-borne Virus Research Center State Key Laboratory of Ecological Pest Control for Fujian and Taiwan CropsFujian Province Key Laboratory of Plant VirologyInstitute of Plant VirologyFujian Agriculture and Forestry UniversityFuzhou 350002China

Tillering is a major determinant of rice plant architecture and grain ***,we report that depletion of rice OsNRPD1a and OsNRPD1b,two orthologs of the largest subunit of RNA polymerase IV,leads to a high-tillering phenotype,in addition to dwarfism and smaller ***1a and OsNRPD1b are required for the production of 24-nt small interfering RNAs that direct DNA methylation at transposable elements(TEs)including miniature inverted-repeat TEs(MITEs).Interestingly,many genes are regulated either positively or negatively by TE *** them,OsMIR156d and OsMIR156j,which promote rice tillering,are repressed by CHH methylation at two MITEs in the *** contrast,D14,which suppresses rice tillering,is activated by CHH methylation at an MITE in its *** findings reveal regulation of rice tillering by RNA-directed DNA methylation at MITEs and provide potential targets for agronomic trait enhancement through epigenome editing.

关键词： RdDM siRNA MITEs OsMIR156 D14 tiller

Effect of wortmannin and 3-AB on G_2 phase arrest and transient failure to activate MPF induced by ionizing irradiation

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Chinese Science Bulletin 1998年第18期43卷 1550-1555页

作者： meng xiangbing and SUN Zhixian Beijing Institute of Radiation Medicine, Beijing 100850, China Beijing Institute of Radiation Medicine Beijing China

Active mitosis promoting factor (MPF) composed of p34 cdc2 with cyclin B1 is required for the cell cycle transition from G 2 to M phase. Recent studies have demonstrated that ionizing radiation (IR) exposures associated with inhibition of p34 cdc2 activity was the mechanism of G 2 arrest. At low dose, IR causes transient failure to dephosphorylate p34 cdc2 instead of suppression of cyclin B or MPF formation of cyclin B with p34 cdc2 complex. But the signaling events that regulate p34 cdc2 in irradiated cells remain unclear. This note demonstrates that PI3 kinase family inhibitor wortmannin (wort) and PARP specific inhibitor 3_AB can reduce the G 2 arrest induced by 2Gy γ_ray. Immunoprecipitation studies demonstrate that wort and 3_AB can facilitate dephosphorylation of p34 cdc2 in G 2 phase arrest induced by radiation. These findings suggest that wort sensitive pathway and PARP may be involved in initiating the signal transduction of G 2 phase arrest caused by IR.

关键词： G 2 phase arrest ionizing radiation MPF wortmannin 3_AB.

Contribution of alternative splicing to breast cancer metastasis

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Journal of Cancer Metastasis and Treatment 2019年第3期4卷 73-79页

作者： xiangbing meng Shujie Yang Jun Zhang Huimin Yu Department of Obstetrics and Gynecology University of Iowa Carver College of MedicineIowa CityIA 52242USA Holden Comprehensive Cancer Center University of Iowa Carver College of MedicineIowa CityIA 52242USA Division of Hematology Oncology and Blood&Marrow TransplantationDepartment of Internal MedicineUniversity of Iowa Carver College of MedicineIowa CityIA 52242USA Department of Pathogenic Biology Shenzhen University School of medicineShenzhen 518060China

Alternative splicing is a major contributor to transcriptome and proteome diversity in eukaryotes. Comparing to normal samples, about 30% more alternative splicing events were recently identified in 32 cancer types included in The Cancer Genome Atlas database. Some alternative splicing isoforms and their encoded proteins contribute to specific cancer hallmarks. In this review, we will discuss recent progress regarding the contributions of alternative splicing to breast cancer metastasis. We plan to dissect the role of MTDH, CD44 and their interaction with other mRNA splicing factors. We believe an in-depth understanding of the mechanism underlying the contribution of splicing to breast cancer metastasis will provide novel strategies to the management of breast cancer.

关键词： Breast cancer metastasis CD44 MTDH splicing epithelial-mesenchymal transition

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

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

Generating broad-spectrum tolerance to ALS-inhibiting herbicides in rice by base editing

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Science China(Life Sciences) 2021年第10期64卷 1624-1633页

作者： Rui Zhang Sha Chen xiangbing meng Zhuangzhuang Chai Delin Wang Yuge Yuan Kunling Chen Linjian Jiang Jiayang Li Caixia Gao State Key Laboratory of Plant Cell and Chromosome Engineering Center for Genome EditingInstitute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing 100049China State Key Laboratory of Plant Genomics Institute of Genetics and Developmental BiologyInnovation Academy for Seed DesignChinese Academy of SciencesBeijing 100101China Key Lab of Pest Monitoring and Green Management MOA/Department of Plant PathologyCollege of Plant ProtectionChina Agricultural UniversityBeijing 100193China

Herbicide-tolerant rice varieties generated by genome editing are highly desirable for weed *** have used a cytosine base editor to create a series of missense mutations in the P171 and/or G628 codons of the acetolactate synthase(ALS)gene to confer herbicide tolerance in *** four different missense mutations in the P171 codon,P171S,P171A,P171Yand P171F,exhibited different patterns of tolerance towards five representative herbicides from five chemical families of ALS *** example,P171S and P171A had lower levels of tolerance than P171Y and P171F to bispyribac but not to the other ***,a novel triple mutant(P171F/G628E/G629S)had the highest tolerance to all five tested *** trials showed that both P171F and P171F/G628E/G629S could potentially be used with *** work illustrates an effective way of using base editing to generate herbicide tolerance in elite rice varieties.

关键词： base editing herbicide tolerance rice acetolactate synthase(ALS)

Construction of a Genome-Wide Mutant Library in Rice Using CRISPR/Cas9

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Molecular Plant 2017年第9期10卷 1238-1241页

作者： meng, xiangbing Yu, Hong Zhang, Yi Zhuang, Fengfeng Song, Xiaoguang Gao, Songsong Gao, Caixia Li, Jiayang Chinese Acad Sci Inst Genet & Dev Biol State Key Lab Plant Genom Beijing 100101 Peoples R China Chinese Acad Sci Inst Genet & Dev Biol Natl Ctr Plant Gene Res Beijing 100101 Peoples R China Chinese Acad Sci Inst Genet & Dev Biol State Key Lab Plant Cell & Chromosome Engn Beijing 100101 Peoples R China Beijing ViewSolid Biotechnol Beijing 102206 Peoples R China Univ Chinese Acad Sci Beijing 100049 Peoples R China

Dear Editor Rice （Oryza sativa L.） is one of the world＇s most important staple crops and a powerful model system for studying monocot spe- cies because of its relatively small genome, rich genomic re- sources, and a highly efficient transformation system. With the completion of rice genome sequencing, the challenge of the post-genomic era is to systematically analyze the functions of all rice genes. Gene knockout is a frequently used and effective strategy for achieving this goal. Thus, generation of large-scale mutants at the whole-genome level is of great value for both func- tional genomics and genetic improvement of rice. Traditionally,

关键词： Construction of a Genome-Wide Mutant Library Rice Using CRISPR

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