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《Chinese Journal of Cellular and Molecular Immunology》 2014-01
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Construction of miR-508-5P lentiviral vector and its targeted effect on S-phase kinase-associated protein-2 gene

BAI Jing;XING Yiwen;FAN Weining;GUO Le;LIU Xin;ZHENG Ximing;XU Guangxian;Institute of Laboratory Medicine,Ningxia Medical University;Medical Experimental Center,General Hospital,Ningxia Medical University;  
Objective To construct a lentiviral vector overexpressing miR-508-5p and verify its targeted regulating effect on S-phase kinase-associated protein-2( SKP2) gene. Methods The stem-loop structure RNA of miR-508-5p was obtained by chemical synthesis and cloned into the linearized pSicoR plasmid. The positive recombinants,which had been identified by restriction enzyme digestion and DNA sequencing method,were transfected into HEK293T cells. Meanwhile,the potential target gene of SKP2-3'UTR,which was complementary with miR-508-5p,was obtained and cloned into the linearized pMIRReport plasmid. The targeted effect of miR-508-5p on SKP2 gene was verified by the relative luciferase activity measurement, Western blotting and real-time PCR. Results The PCR,restriction enzyme digestion and DNA sequencing analysis demonstrated that the recombinant plasmids of pSicoR-miR-508-5p and pMIR-Report-SKP2 3'-UTR were constructed successfully. And it was showed that over-expression of miR-508-5p suppressed the mRNA and protein expression level of SKP2 significantly( P 0. 05); and suppression of miR-508-5p expression increased the mRNA and protein expression level of SKP2 significantly( P 0. 05). Conclusion We have successfully constructed the lentiviral vector containing miR-508-5p gene. MiR-508-5p can suppress SKP2 gene expression by targeting the specific sequence of SKP2-3'UTR.
【Fund】: 国家自然科学基金(31160494);; 教育部“新世纪优秀人才支持计划”(NCET 11-11023);; 中国博士后特别基金(2012T50834);; 中国博士后基金(20110491554)
【CateGory Index】: R346
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 TANG Wei-jun*,ZHANG Xing-ping,ZOU Hong-yuan,PENG Chun-fang(*Department of Oncology,Central Hospital of Jiangjin District,Chongqing City,Chongqing 402260,China);Effect of down-regulation of Skp2 gene expression on biological characters of HepG2 cells as well as expressions of SP1 and p27 proteins[J];Chinese Journal of Biologicals;2012-01
【Co-citations】
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1 Yan-chang Sun;Cheng-cheng Guo;Ke Sai;Jian Wang;Jie Wang;Fu-rong Chen;Zong-ping Zhang;Zhong-ping Chen;Guangdong Medical College;Department of Neurosurgery ,Guangdong Medical University Zhanjiang Cancer Hospital;Department of Neurosurgery, Sun Yat-sen University Cancer Center,Guangzhou/State Key Laboratory of Oncology in South China;;miR-181b Enhances Sensitivity to Teniposide in Glioma Cell Line U87[J];Chinese Journal of Neuro-Oncology;2013-02
2 WANG Zhang - xun;ZHOU Quan;MENG Hui - ming;ZHANG Jun;HUANG Bo;School of Plant Protection,Anhui Agricultural University;Anhui Provincial Key Laboratory of Microbial Pest Control,Anhui Agricultural University;;Differential Expression Analysis of MicroRNA in Cicada Slough and Hemolymph Induction Conditions from Metarhizium anisopliae[J];Journal of Anhui Science and Technology University;2013-05
3 Wenwen Jia;Wen Chen;Jiuhong Kang;Clinical and Translational Research Center of Shanghai First Maternity and Infant Health Hospital,Shanghai Key Laboratory of Signaling and Disease Research,School of Life Science and Technology,Tongji University;;The Functions of MicroRNAs and Long Non-coding RNAs in Embryonic and Induced Pluripotent Stem Cells[J];基因组蛋白质组与生物信息学报(英文版);2013-05
4 Chen Juanjuan;Ai Jiangang;Huang Shifeng;Cao Ju;Zhang Liping;Department of Clinical Laboratory,First Affiliated Hospital,Chongqing Medical University;;HBx protein-induced up-regulation of miR-221 promotes HepG2 cell malignant proliferation by targeting ERα[J];Journal of Third Military Medical University;2013-20
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6 Juan Liu;Chang-Ping Wu;Bin-Feng Lu;Jing-Ting Jiang;Department of Tumor Biological Treatment,The Third Affiliated Hospital of Soochow University;Department of Immunology,University of Pittsburgh School of Medicine;;Mechanism of T cell regulation by microRNAs[J];癌症生物学与医学(英文版);2013-03
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1 Ruiwen Fan Shanshan Yang Zhanquan Shi Kaiyuan Ji Xiaoyun Jia Jianbo Yao George W Smith Changsheng Dong~*;Identification of a novel miRNA important for melanogenesis in alpaca(Lama Pacos)[A];[C];2013
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1 LUO Xin,LIU Dan-dan,QI Hong-bo,YAO Zhen-wei,CHEN Guo-qing(Department of Obstetrics and Gynecology,First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,China);Effect of shRNA Lentiviral Vector Targeting Human Growth Arrest and DNA Damage 45α Gene on Disfunction of Human Umbilical Vein Endothelial Cells Due to Hypoxia Stress[J];Chinese Journal of Biologicals;2010-11
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