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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 | 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 |
| 5 | 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 |
| 6 | 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 |
| 7 | Differences in functional properties and biochemical characteristics of congeneric rice proteins显示文摘 | Xiaohong Cao Huanbin Wen Cuijuan Li | 2009 | Journal of Cereal Science2009，50，： | 1 |
| 8 | 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 |
| 9 | Long-range ordering of composites for organic electronics:TIPS-pentacene single crystals with incorporated nano-fibers显示文摘Multi-component active materials are widely used for organic electronic devices, with every component contributing complementary and synergistic optoelectronic functions. Mixing these components generally leads to lowered crystallinity and weakened charge transport. Therefore, preparing the active materials without substantially disrupting the crystalline lattice is highly desired. Here, we show that crystallization of TIPS-pentacene from solutions in the presence of fluorescent nanofibers of a perylene bisimide derivative(PBI) leads to formation of composites with nanofiber guest incorporated in the crystal host. In spite of the binary composite structure, the TIPS-pentacene maintains the singlecrystalline nature. As a result, the incorporation of the PBI guest introduces additional fluorescence function but does not significantly reduce the charge transport property of the TIPS-pentacene host,exhibiting field-effect mobility as high as 3.34 cm^2 V^(-1) s^(-1) even though 26.4% of the channel area is taken over by the guest. As such, this work provides a facile approach toward high-performance multifunctional organic electronic materials. | Huanbin Li Guobiao Xue Jiake Wu Wenqiang Zhang Zhuoting Huang Zengqi Xie Huolin L.Xin Gang Wu Hongzheng Chen Hanying Li | 2017 | Chinese Chemical Letters2017，28，11： | 0 |
| 10 | ASAP3 regulates microvilli structure in parietal cells and presents intervention target for gastric acidity显示文摘Gastric acidity-associated disorders such as peptic ulcer and reflux diseases are widespread,and the reported resistance and side effects of currently used medicines suggest an urgent requirement for alternative therapeutic approaches.Here we demonstrate a critical role of ASAP3 in regulating the microvilli structure of parietal cells in vivo,and reveal the feasibility of controlling gastric acidity by targeting ASAP3.Conditional knockout of ASAP3 in mice caused elongation and stacking of microvilli in parietal cells,and substantially decreased gastric acid secretion.These were associated with active assembly of F-actin caused by a higher level of GTP-bound Arf6 GTPase.Consistently,a small molecular compound QS11 inhibited ASAP3 function and significantly reduced gastric acidity in vivo.Of note,the expression of ASAP3 was positively correlated with gastric acid secretion in 90 human cases,and high expression of ASAP3 was associated with reflux disease and peptic ulcer.These results reveal for the first time that ASAP3 regulates the microvilli structures in parietal cells.Our data also suggest ASAP3 as a feasible and drugable therapeutic target for gastric acidity-associated diseases. | Jin Qian Yueyuan Li Han Yao Haiying Tian Huanbin Wang Luoyan Ai Yuanhong Xie Yujie Bao Lunxi Liang Ye Hu Yao Zhang Jilin Wang Chushu Li Jiayin Tang Yingxuan Chen Jie Xu Jing-Yuan Fang | 2017 | Signal Transduction and Targeted Therapy2017，2，1： | 0 |
| 11 | 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 |
| 12 | 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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