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| 1 | Improved Base Editor for Efficiently Inducing Genetic Variations in Rice with CRISPR/Cas9- Guided Hyperactive hAID Mutant显示文摘 | Bin Ren Fang Yan Yongjie Kuang Na Li Dawei Zhang Xueping Zhou Honghui Lin Huanbin Zhou | 2018 | Molecular Plant2018，11，4： | 46 |
| 2 | Base-Editing-Mediated Artificial Evolution of OsALS1 In Planta to Develop Novel Herbicide-Tolerant Rice Germplasms显示文摘Recently developed CRISPR-mediated base editors,which enable the generation of num erous nucleotide changes in target genomic regions,have been widely adopted for gene correction and generation of crop germ plasms containing im portant gain-of-function genetic variations.How ever,to engineer target genes with unknown functional SNPs remains challenging.To address this issue,we present here abase-e diting-mediated gene evolution(BEMGE)m ethod,employing both Cas9n-based cytosine and adenine base editors as well as a single-guide RNA(sgRNA)library tiling the full-length coding region,for developing novel rice germ plasm swith mutations in any endogenous gene.To this end,OsALS1 was artificially evolved in rice cells using BEMGE through both Agrobacterium-mediated and particle-bom bardment-mediated transform ation.Four different types of amino acid substitutions in the evolved OsALS1,derived from two sites that have never been targeted by natural or human selection during rice dom estication,were identified,conferring varying levels of tolerance to the herbicide bispyribac-sodium.Furtherm ore,the P171F substitution identified in a strong OsALS1 allele was quickly introduced into the commercial rice cultivar Nangeng 46 through precise base editing w ith the corresponding base editor and sgRNA.Collectively,these data indicate great potential of BEMGE in creating important genetic variants of target genes for crop improvement. | Yongjie Kuang Shaofang Li Bin Ren Fang Yan Carl Spetz Xiangju Li Xueping Zhou Huanbin Zhou | 2020 | Molecular Plant2020，13，4： | 37 |
| 3 | A CRISPR/Cas9 toolkit for efficient targeted base editing to induce genetic variations in rice显示文摘Dear Editor,Gene-targeting technologies using sequence specific nucleases have been widely adopted to induce genetic modifications in both plant molecular biology research and crop improvement,of which generating targeted point mutation for gain-of-function phenotype is of great value.Although tremendous efforts have been made in developing geneinsertion or replacement approaches by | Bin Ren Fang Yan Yongjie Kuang Na Li Dawei Zhang Honghui Lin Huanbin Zhou | 2017 | Science China(Life Sciences)2017，60，5： | 28 |
| 4 | Highly Efficient A. T to G. C Base Editing by Cas9n- 3uided tRNA Adenosine Deaminase in Rice显示文摘 | Fang Yan Yongjie Kuang Bin Ren Jingwen Wang Dawei Zhang Honghui Lin Bing Yang Xueping Zhou Huanbin Zhou | 2018 | Molecular Plant2018，11，4： | 26 |
| 5 | Cas9-NG Greatly Expands the Targeting Scope of the Genome-Editing Toolkit by Recognizing NG and Other Atypical PAMs in Rice显示文摘CRISPR technologies enabling precise genome manipulation are valuable for gene function studies and molecular crop breeding. However, the requirement of a protospacer adjacent motif (PAM)y such as NGG and TTN, for Cas protein recognition restricts the selection of targetable genomic loci in practical applications of CRISPR technologies. Recently Cas9-NG, which recognizes a minimal NG PAM, was reported to expand the targeting space of genome editing in human cells, but it remains unclear whether this Cas9 variant can be used in plants. In this study, we evaluated the nuclease activity of Cas9-NG toward various NGN PAMs by targeting endogenous genes in transgenic rice. We found that Cas9-NG edits all NGG, NGA, NGT, and NGC sites with impaired activity, while the gene-edited plants were dominated by monoallelic mutations. Cas9-NG-engineered base editors were then developed and used to generate O s B Z R I gainof- function plants that can not be created by other available Cas9-engineered base editors. Moreover, we showed that a Cas9-NG-based transcriptional activator efficiently upregulated the expression of endogenous target genes in rice. In addition, we discovered that Cas9-NG recognizes NAC, NTG, NTT, and NCG apart from NG PAM. Together, these findings demonstrate that Cas9-NG can greatly expand the targeting scope of genome-editing tools, showing great potential for targeted genome editing, base editing, and genome regulation in plants. | Bin Ren Lang Liu Shaofang Li Yongjie Kuang Jingwen Wang Dawei Zhang Xueping Zhou Honghui Lin Huanbin Zhou | 2019 | Molecular Plant2019，12，7： | 18 |
| 6 | Synergistic immunoreaction of acupuncture-like dissolving microneedles containing thymopentin at acupoints in immune-suppressed rats显示文摘Dissolving microneedles carried drug molecules can effectively penetrate the stratum corneum of skin to improve the transdermal drug delivery. The traditional Chinese medicine acupuncture is based on the needle stimulation at a specific location(acupoint) to generate and transmit biochemical and physiological signals which alter the pathophysiological state of patients. However, the pain associated with conventional acupuncture needles and the requirement of highly trained professionals limit the development of acupuncture in non-Asian countries. The purpose of this study is to investigate whether the dissolving microneedles can be utilized as a self-administered painless replacement for acupuncture and locally released drug molecules can achieve expected therapeutic outcomes. Immunosuppressive rats were treated with acupuncture at Zusanli(ST36) acupoint using microneedles containing thymopentin.The immune functions and psychological mood of the immunosuppressed animals were examined. The proliferation of splenocytes was examined by CCK-8 assay. CD4 and CD8 expression patterns in spleen cells were detected by flow cytometry. The current study showed that use of either microneedles containing thymopentin or conventional acupuncture both resulted in immune cell proliferation, which was confirmed by flow cytometry. Furthermore, either conventional acupuncture or microneedles were able to effectively mitigate the anxiety caused by immune-suppression when applied on the ST36. | Qian Zhang Chuncao Xu Shiqi Lin Huanbin Zhou Gangtao Yao Hu Liu Lili Wang Xin Pan Guilan Quan Chuanbin Wu | 2018 | Acta Pharmaceutica Sinica B2018，8，3： | 9 |
| 7 | High-efficiency and multiplex adenine base editing in plants using new TadA variants显示文摘Recently reported adenine base editors(ABEs)exhibit powerful potential for targeted gene correction as well as developing gain-of-function mutants and novel germplasms for both gene function studies and crop breeding.However,editing efficiency varies significantly among different target sites.Here,we investigated the activities of three evolved E.coli adenosine deaminase TadA variants(TadA8e,TadA8.17,and TadA8.20)side-by-side in transgenic rice.We found that TadA8e outperforms TadA8.17 and TadA8.20,and induces efficient A-to-G conversion at all tested sites in the rice genome,including those that were un-editable by ABE7.10 in our previous experiments.Furthermore,V82S/Q154R mutations were incorporated into TadA8e,resulting in a new variant that we named TadA9.Our data show that TadA9 is broadly compatible with CRISPR/SpCas9,CRISPR/SpCas9-NG,and CRISPR/SpRY,as well as CRISPR/ScCas9 nickase systems,achieving comparable or enhanced editing in a larger editing window at diverse PAM sites as compared with TadA8e.Finally,TadA9 was used to simultaneously install novel SNPs in four endogenous herbicide target genes in the commercial rice cultivar Nangeng 46 for potential field application in.weed control.Collectively,we successfully generated a series of novel ABEs that can efficiently edit adenosines in the rice genome.Our findings suggest that TadA9 and TadA8e have great potentials in the development of plant base editors and crop molecular breeding. | Daqi Yan Bin Ren Lang Liu Fang Yan Shaofang Li Guirong Wang Wenxian Sun Xueping Zhou Huanbin Zhou | 2021 | Molecular Plant2021，14，5： | 8 |
| 8 | Rock paper scissors: CRISPR/Cas9-mediated interference with geminiviruses in plants显示文摘Geminiviruses are a group of circular single-stranded DNA viruses that constitute the largest family of plant viruses. Many diseases resulting from geminivirus infections, such as maize streak disease, cassava mosaic disease, tomato yellow leaf curl disease and cotton leaf curl diseases, cause significant problems in terms of economic losses, posing a serious threat to global crop productivity (Hanley-Bowdoin et al., 2013;Yang et al., 2019). As obligate intracellular parasites, geminiviruses cannot be controlled chemically. Conventional strategies to control geminiviruses rely heavily on the application of pesticides to limit insect vector populations, but it causes environmental concerns. To date, developing novel crop germplasms with durable disease resistance to geminiviruses is still the best strategy to manage viral infections due to its environmental friendliness. However, traditional breeding programs are time-intensive and labor-consuming processes, and transgene-based modern approaches by overexpressing the geminiviral proteins (e.g., the truncated form of the replication initiator protein) failed to confer resistance to geminiviruses due to activated virusinduced transgene silencing. | Xiuling Yang Huanbin Zhou Xueping Zhou | 2019 | Science China(Life Sciences)2019，62，10： | 4 |
| 9 | Conferring Resistance to Plant RNA Viruses with the CRISPR/CasRx System显示文摘Dear Editor,Clustered regularly interspaced short palindromic repeat(CRISPR)-associated system(Cas)is an adaptive immune system discovered in prokaryotic bacteria or archaea that can fend off invading nucleic acids.Because of its simplicity,high efficiency and versatility,CRISPR/Cas system-mediated genome editing has been widely applied in plant research and agricultural production. | Yongsen Cao Huanbin Zhou Xueping Zhou Fangfang Li | 2021 | Virologica Sinica2021，36，4： | 2 |
| 10 | The rice codebook: From reading to editing显示文摘Rice(Oryza sativa)is the main staple food crop that feeds more than half of the world's population.Given its relatively small genome size,abundant genetic resources,and highly efficient transformation technology,rice also remains as an excellent model system for monocot crops.In 2002,the International Rice Genome Sequencing Project(IRGSP)released the draft genome sequence of rice variety Nipponbare,the first crop genetic codebook deciphered right after the human genome(Yu et al.,2002).Since then,rice genome annotation,large-scale genome resequencing of diverse rice germplasm accessions,a comprehensive map of rice genome variations,and high-quality reference genome sequence,especially the gap-free reference genome,were completed successively in the next two decades along with the development of high-throughput DNA sequencing techniques(Wang et al.,2018;Li et al.,2021;Song et al.,2021).Consequently,rice functional genomics research has boomed rapidly,greatly reshaping our knowledge of plant molecular biology and designed crop breeding. | Kejian Wang Huanbin Zhou Qian Qian | 2022 | Molecular Plant2022，15，4： | 1 |
| 11 | Co-evolved plant and blast fungus ascorbate oxidases orchestrate the redox state of host apoplast to modulate rice immunity显示文摘Apoplastic ascorbate oxidases(AOs)play a critical role in reactive oxygen species(RoS)-mediated innate host immunity by regulating the apoplast redox state.To date,little is known about how apoplastic effectors of the riceblast fungus Magnaportheoryzaemodulate the apoplast redox state of rice to subvert plant immunity.In this study,we demonstrated that M.oryzae MoAo1 is an Ao that plays a role in virulence by modulating the apoplast redox status of rice cells.We showed that MoAo1 inhibits the activity of rice OsAO3and OsAO4,which also regulate the apoplast redox status and plant immunity.In addition,we found that MoAo1,OsAO3,andOsAO4 allexhibit polymorphic variations whosevaried interactions orchestrate pathogen virulence and rice immunity.Taken together,our results reveal a critical role for extracellular redox enzymes during rice blast infection and shed light on the importance of the apoplast redox state anditsregulation inplant-pathogeninteractions. | Jiexiong Hu Muxing Liu Ao Zhang Ying Dai Weizhong Chen Fang Chen Wenya Wang Danyu Shen Mary Jeanie Telebanco-Yanoria Bin Ren Haifeng Zhang Huanbin Zhou Bo Zhou Ping Wang Zhengguang Zhang | 2022 | Molecular Plant2022，15，8： | 0 |
| 12 | CRISPR/Sc^(++)-mediated genome editing in rice显示文摘Streptococcus canis Cas9(ScCas9)is an RNAguided endonuclease with NNG protospacer adjacent motif(PAM)specificity whose genomeediting activity in rice is locus-dependent.Here we investigated the performance of a ScCas9 variant named Sc^(++)at different NNG PAM sites in the rice genome;Sc^(++)harbors a T1227K mutation and the substitution of a positively charged loop(residues 367-376).Sc^(++)nuclease achieved broader genome editing compared to the original ScCas9,and its nickase improved targeted base editing in transgenic rice plants.Using the high-efficiency adenine base editor rBE73b,we generated many new OsGS1 alleles suitable for screening of rice germplasm for potential herbicide resistance in the future.The CRISPR/Sc^(++)system expands the genomeediting toolkit for rice. | Guigen Ma Yongjie Kuang Zhenwan Lu Xueqi Li Ziyan Xu Bin Ren Xueping Zhou Huanbin Zhou | 2021 | Journal of Integrative Plant Biology2021，63，9： | 0 |
| 13 | Simultaneous quadratic performance stabilization for linear time-delay systems显示文摘A newly designed approach of simultaneous stabilization is given for linear discrete time-delay systems. The problem of stabilization for a collection of systems is discussed initially. Adequate condition are obtained in terms of linear matrix inequalities (LMIs) which are independent of time delays such that the resultant collection of discrete time-delay systems are stable with an upper bound of the quadratic performance index. Subsequently, controllers are designed such that the resultant closed-loop discrete time-delay systems are simultaneously stabilized with the upper bound of the quadratic performance index. Finally,a numerical example is given to illustrate the design method. | Chen Yuepeng Zhou Zude Liu Huanbin Zhang Qingling | 2006 | Journal of Systems Engineering and Electronics2006，17，4： | 0 |
| 14 | Three novel alleles of OsGS1 developed by base-editing-mediated artificial evolution confer glufosinate tolerance in rice显示文摘Only few glufosinate-tolerant genes,such as phosphinothricin acetyltransferase(PAT)and bialaphos resistance(bar)identified from Streptomyces,are currently available for developing genetically modified rice in agricultural application.Following the rapid development of genome editing technology,generation of novel glufosinate-tolerant gene resources through artificial evolution of endogenous genes is more promising and highly desirable in rice molecular breeding program.In this study,the endogenous Glutamine synthetase1(OsGS1)was artificially evolved by base-editing-mediated gene evolution(BEMGE)in rice cells to create novel alleles conferring glufosinate tolerance in rice germplasms.Two novel glufosinate-tolerant OsGS1 alleles(OsGS1-AVPS and OsGS1-+AF)and one reported tolerant allele(OsGS1-SGTA)were successfully identified from approximately 4200 independent hygromycin-tolerant calli.Germination assays and spray tests revealed that these three OsGS1 alleles confer glufosinate tolerance in rice.Furthermore,OsGS1-AVPS and OsGS1-SGTA were quickly deployed into the elite rice cultivar Nangeng 46 through precise base editing.Overall,our results demonstrate the feasibility of developing glufosinate-tolerant rice by editing an endogenous rice gene in molecular breeding programs. | Bin Ren Yongjie Kuang Ziyan Xu Xuemei Wu Dawei Zhang Fang Yan Xiangju Li Xueping Zhou Guirong Wang Huanbin Zhou | 2023 | The Crop Journal2023，11，2： | 0 |
| 15 | Adenine base editor incorporating the N-methylpurine DNA glycosylase MPGv3 enables efficient A-to-K base editing in rice显示文摘Dear Editor,Crop genetic diversity and elite agronomic traits are mainly caused by genetic variants,approximately half of which are single-nucleotide polymorphisms.Precise nucleotide substitution through CRISPR–Cas-mediated base editors has been widely used to correct defective alleles and create novel alleles by artificial evolution for rapid crop genetic improvement.Since 2017,cytosine base editors(CBEs)and adenine base editors(ABEs)have been successively developed in many plant species and have been continuously optimized to generate highly efficient C-to-T and A-to-G transitions,as well as by-product C-to-A/G conversion with low efficiency(Ren et al.,2018;Yan et al.,2021). | Xuemei Wu Bin Ren Lang Liu Shengqun Qiu Xin’ge Li Peijing Li Fang Yan Honghui Lin Xueping Zhou Dawei Zhang Huanbin Zhou | 2023 | Plant Communications2023，4，6： | 0 |
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