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Inhibition of genes expression of SARS coronavirus by synthetic small interfering RNAs

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cell Research 2005年第3期15卷 193-200页

作者： yi shi,De Hua YANG,Jie XIONG,Jie JIA,Bing HUANG,You Xin jin* state key laboratory of molecular biology,institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences,shanghai 200031,china statekeylaboratoryofmolecularbiology InstituteofBiochemistryandCellBiologyShanghaiInstitutesforBiologicalSciencesChineseAcademyofSciencesShanghai200031China

RNA interference (RNAi) is triggered by the presence of a double-stranded RNA (dsRNA), and results in the silencing of homologous gene expression through the specific degradation of an mRNA containing the same sequence. dsRNA- mediated RNAi can be used in a wide variety of eucaryotes to induce the sequence-specific inhibition of gene expression. Synthetic 21-23 nucleotide (nt) small interfering RNA (siRNA) with 2 nt 3’ overhangs was recently found to mediate efficient sequence-specific mRNA degradation in mammalian cells. Here, we studied the effects of synthetic siRNA duplexes targeted to SARS coronavirus structural proteins E, M, and N in a cell culture system. Among total 26 siRNA duplexes, we obtained 3 siRNA duplexes which could sequence-specifically reduce target genes expression over 80% at the concentration of 60 nM in Vero E6 cells. The downregulation effect was in correlation with the concentrations of the siRNA duplexes in a range of 0~60 nM. Our results also showed that many inactive siRNA duplexes may be brought to life simply by unpairing the 5’ end of the antisense strands. Results suggest that siRNA is capable of inhibiting SARS coronavirus genes expression and thus may be a new therapeutic strategy for treatment of SARS.

关键词： SARS small interfering RNA Vero E6 cells EGFP fusion protein antiviral therapy.

Dose-dependent Inhibition of Gynecophoral Canal Protein Gene Expression in Vitro in the Schistosome(Schistosomajaponicum)by RNA Interference

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Acta Biochimica et Biophysica Sinica 2005年第6期37卷 386-390页

作者： Guo-Feng CHENG~1 Jiao-Jiao LIN~1 yi shi~2 You-Xin jin~2 Zhi-Qiang FU~1 Ya-Mei jin~1 Yuan-Cong ZHOU~2 You-Min CA~1 1 shanghai institute of Animal Parasitology, chinese academy of Agricultural sciences, key Lahoratory of Animal Parasitology,Ministry of Agriculture, shanghai 200232, china 2 state key laboratory, of molecular biology institute of biochemistry and cell biology shanghai institutes for biological sciences, chinese academy of sciences. shanghai 200031, china shanghai institute of Animal Parasitology Chinese Academy of Agricultural Sciences Key Laboratory of Animal Parasitology Ministry of Agriculture Shanghai 200232 China.

The gynecophoral canal protein gene SjGCP of Schistosoma japonicum that is necessary forthe pairing between the male and female worms is specifically expressed in the adult male *** is widely distributed in the adult female worm after *** transcription-polymerase chainreaction(RT-PCR)and immunofluorescence were employed to analyze the relationship between the RNAieffect and dsRNA dosage in the *** results revealed that the inhibition of SjGCP expression bysiRNA is ***-PCR analysis showed that the SjGCP transcript level was reduced by 75%when 100 nM dsRNA was applied.

关键词： RNA interference gynecophoral canal protein gene Schistosoma japonicum dosage

A method for microRNA targets identification based on RISC complex co-immuno-precipitation

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A method for microRNA targets identification based on RISC c...

华东六省一市生物化学与分子生物学会2008年学术交流会

作者： yi shi Botao Zhao Bing Huang Yang Li Wenmin Qin Chenguo Yao youxin jin state key laboratory of molecular biology,shanghai institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences

MicroRNAs(miRNAs)are a large class of noneoding RNAs that play a pivotal role in gene *** of miRNAs have been discovered in higher eukaryotes and many of them exhibit different roles in development,differentiation,proliferation,apoptosis,metabolism, and tumor initiation and progression *** until now,only a few miRNA downstream target genes have been *** we developed a method combined RNA-induced silencing complex(RISC)purification and bound mRNA’s cloning by PCR to identify specific miRNA’s target ***,we used Ago 2 antibody to co-immune-precipitate RISC in Hela *** the bound mRNAs were extracted and the complementary cDNA strands were sythesized by reverse transcription *** fragments between the miRNA binding sites and the 3’end of mRNAs could be amplified by PCR and cloned into T vector for identification by *** using this method,we identified 21 miR-16’s target genes from 65 clones in Hela cells(and these target genes were validated by reporter assay).In conclusion,we devised an efficient and low cost method for miRNA target genes identificaiton and it will be a powerful tool in miRNA function research.

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The polarity of E153 in *** TrpRS is essential for the activation of tryptophan

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The polarity of E153 in *** TrpRS is essential for the activ...

第三届全国暨2003年中外RNA研讨会

作者： Jie Jia, Xianglong Chen, yi shi and You-xin jin state key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai institutes for biologicalsciences, chinese academy of sciences, shanghai 200031, P. R. china

From residues P143 to residues A155 in *** TrpRS is a very conserved domain located between "GXDQ" and "KMSKS" in the tryptophanyl-tRNA synthetases. These 13 amino acids belong to the catalytic domain that charges for the activation of tryptophan and the aminoacylation of tRNA. Based on the recent data of the TrpRS-TAM complex crystal structure, the position of residues and the interaction between the residues and tryptophan or ATP have been illustrated in detail. E153 locates in the binding pocket of tryptophan. It provides an H-bond to or an electrostatic interaction with the substrate-tryptophan. To study the importance of glutamate, E153 was mutated to D153, K153 and G153, respectively. The kinetics parameters of three mutant TrpRSs were determined. There were no significant changes in the values of Km and kcal when El 53 was mutated to D153. After El 53 became to K153, both the Km and the kcal increased near 30-fold. But the relative activity of the second mutant not changed so much compared with the wild-type TrpRS. Once E153 was changed to G153, the reaction of tryptophan activation seemed to not occur even as the concentration of TrpRS had been added to 10,000-fold than the other usage. The CD spectra of the wild-type TrpRS and three mutants were similar;the mutagenesis had little effect on the secondary structure of the enzymes. Both the kinetics data and structural parameters support that the polarity of E153 is important to the processing of tryptophan activation.

关键词： TrpRS polarity of E153 activation of tryptophan

A Method for MicroRNA Targets Identification Based on RISC Complex Coimmunoprecipitaion

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A Method for MicroRNA Targets Identification Based on RISC C...

2008年全国生物化学与分子生物学学术大会

作者： yi shi Bo-tao Zhao Bing Huang Yang Li Wen-min Qin Chen-guo Yao You-xin jin (state key laboratory of molecular biology,shanghai institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences.shanghai 200031)

MicroRNAs (miRNAs) are a large class of noncoding RNAs that play a pivotal role in gene regulation Hundreds of miRNAs have been discoved in higher eukaryotes and many of them exhibit different roles in development,dif...

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Establishment of a Screening System for Selection of siRNA Target Sites Directed against Hepatitis B Virus Surface Gene

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Acta Biochimica et Biophysica Sinica 2005年第5期37卷 310-316页

作者： Xiu-Min ZHOU~(1.3) Ju-Sheng LIN~1 yi shi~3 De-An TIAN~2 Huan-Jun HUANG~2 He-Jun ZHOU~1 You-Xin jin~3 1 institute of Liver Diseases. Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, china: 2 Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, china 3 state key laboratory of molecular biology, institute of biochemistry and cell biology,shanghai institutes for biological sciences,chinese academy of sciences, shanghai 200031, china instituteofLiverDiseases TongjiHospitalTongjiMedicalCollegeHuazhongUniversityofScienceandTechnologyWuhan4

To study the inhibitory effects of plasmid-derived small interfering RNA(siRNA)andsynthetic siRNA on the expression of the hepatitis B virus surface(HBs)gene,three plasmid-derived siRNAsand one synthetic siRNA that complement the coding region of the HBs gene were *** HBsexpression plasmid pHBs-EGFP was also *** cells were co-transfected with pHBs-EGFPand the above *** HBs mRNA quantities were measured by reverse-transcription PCR,and thelevel of HBs-EGFP fusion protein was quantified by fluorescent *** concentrations of thehepatitis B virus surface antigen(HBsAg)derived from the culture supernatant of transfected HepG2.2.15cells were measured by an enzyme-linked immunosorbent assay(ELISA)*** results showed that thethree plasmid-derived siRNAs and the synthetic siRNA can effectively reduce the quantities of HBs mRNAand *** plasmid-derived siRNA psiRNA1 was found to be the most effective inhibitor of *** can inhibit HBs-EGFP expression by 63.3% and suppress HBs mRNA by 75.6%.To furthersubstantiate the above observations,psiRNA1 was transfected into HepG2.2.15 cells(an HBV secreting cellline).The transfections resulted in almost complete blockage of HBsAg production,whereas control vector-transfected cells secreted high levels of HBsAg 7 days *** conclusion,our data suggeststhat RNA interference(RNAi)is an efficient approach for reducing the level of HBs transcripts and proteinsand for suppressing HBsAg production.

关键词： RNA interference(RNAi) small interference RNA(siRNA) hepatitis B virus surface gene(HBs gene)

Androgen activates PEG10 to promote carcinogenesis in hepatic cancer cells Running title: Hepatocarcinogenesis and PEG10 activation

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Androgen activates PEG10 to promote carcinogenesis in hepati...

中国免疫学会第五届全国代表大会暨学术会议

作者： Xiong Jie, Li Chaoqun, Yang Dehua, shi yi, Hu Chunsong, Gong Feili, jin Boquan, jin youxin, Tan jinquan (Department of Immunology, and laboratory of Allergy and Clinical Immunology, institute of Allergy and Immune-related Diseases and Medical Research Center, Wuhan University School of Medicine, Wuhan, 430071 The state key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of Science, shanghai, 200031 Department of Infectious Diseases, Ruijin Hospital, shanghai Second Medical University. shanghai, 200025 College of Life sciences, South china Normal University, Guangzhou, 510631 Department of Immunology, College of Basic Medical sciences, Anhui Medical University, Hefei 230032 Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan Department of Immunology, Fourth Military Medical University, Xian, P. R. china

The molecular mechanisms of hepatocellular carcinoma (HCC) with a striking higher prevalence in male remain to be fully elucidated. We here report the differentially expression of androgen receptor (AR) in different HCC cells and normal hepatocytes. AR agonist dihydrotestosterone (DHT) can promote HCC cell growth and apop-totic resistance, whereas AR antagonist can significantly block the effects of DHT on the HCC cells. Paternally Ex-

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OsNLP3 enhances grain weight and reduces grain chalkiness in rice

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Plant Communications 2024年第10期5卷 82-99页

作者： Liang-Qi Sun Yu Bai Jie Wu shi-Jun Fan Si-Yan Chen Zheng-yi Zhang jin-Qiu Xia shi-Mei Wang Yu-Ping Wang Peng Qin shi-Gui Li Ping Xu Zhong Zhao ChengBin Xiang Zi-Sheng Zhang Division of Life sciences and Medicine Division of Molecular&Cell BiophysicsHefei National Science Center for Interdisciplinary Sciences at the MicroscaleUniversity of Science and Technology of ChinaThe Innovation Academy of Seed DesignChinese Academy of SciencesHefeiAnhui Province 230027China Experimental Center of Engineering and Materials Science University of Science and Technology of ChinaHefei 230027China Rice Research institute Anhui Academy of Agricultural ScienceHefeiChina state key laboratory of Crop Gene Exploration and Utilization in Southwest china Rice Research InstituteSichuan Agricultural UniversityChengduSichuanChina shanghai key laboratory of Plant Functional Genomics and Resources Shanghai Chenshan Botanical GardenShanghai Chenshan Plant Science Research CenterChinese Academy of SciencesShanghaiChina

Grain weight,a key determinant of yield in rice(Oryza sativa L.),is governed primarily by genetic factors,whereas grain chalkiness,a detriment to grain quality,is intertwined with environmental factors such as mineral ***(N)is recognized for its effect on grain chalkiness,but the underlying molecular mechanisms remain to be *** study revealed the pivotal role of rice NODULE INCEPTION-LIKE PROTEIN 3(OsNLP3)in simultaneously regulating grain weight and grain *** investigation showed that loss of OsNLP3 leads to a reduction in both grain weight and dimension,in contrast to the enhancement observed with OsNLP3 ***3 directly suppresses the expression of OsCEP6.1 and OsNF-YA8,which were identified as negative regulators associated with grain ***,two novel regulatory modules,OsNLP3-OsCEP6.1 and OsNLP3-OsNF-YA8,were identified as key players in grain weight ***,the OsNLP3-OsNF-YA8 module not only increases grain weight but also mitigates grain chalkiness in response to *** research clarifies the molecular mechanisms that orchestrate grain weight through the OsNLP3-OsCEP6.1 and OsNLP3-OsNF-YA8 modules,highlighting the pivotal role of the OsNLP3-OsNF-YA8 module in alleviating grain *** findings reveal potential targets for simultaneous enhancement of rice yield and quality.

关键词： OsNLP3 OsCEP6.1 OsNF-YA8 grain weight grain chalkiness rice Oryza sativa L.

RNA-interference-based functional research on rat epididymis-specific gene 16 and human paternally expressed gene PEG10 and the therapeutic applications of RNA interference in HBV,SARS,liver cancer,liver fibrosis and keloid fibroblast

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RNA-interference-based functional research on rat epididymis...

第四届全国RNA进展研讨会

作者： shi Y, Xiong J, Zhou XM, Yang DH, Li GM, Gao ZY and jin YX The state key laboratory of molecular biology, institute of biochemistry and cell biology, shanghai institutes for biological sciences, chinese academy of Science, shanghai 200031, china. Department of Immunology, Wuhan University School of Medicine, Wuhan, Hubei 430071, china. institute of Liver Diseases, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, china.College of Life Science, South china Normal University, Guangzhou 510631, china. Department of Gastroenterology, Xinhua Hospital, shanghai Second Medical University, shanghai 200092, china. Department of Plastic Surgery, Changzheng Hospital, Second Military Medical University, shanghai 200003, china.

RNA interference or RNAi is a powerful cellular mechanism of eukaryotic gene regulation in which a short sequence of double-stranded RNA (dsRNA) specifically down-regulates expression of the associated gene. In mammalian cells, siRNAs do not activate interferon pathway genes, thereby making these powerful tools for basic and applied researches. Our application of RNAi ranged from the study of gene function to therapy of diseases. We have designed siRNA targeting the rat epididymis-specific gene 16 and the human paternally expressed gene PEG10 to explore their function.

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