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《China Biotechnology》 2015-05
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The Establishment of the Methodology used for In Situ Detecting Lentiviral Vector Transcriptional Read-through

GAO Yue;TAN Shuo;REN Zhao-rui;ZHANG Jing-zhi;Shanghai Institute of Medical Genetics,Children's Hospital of Shanghai.Children's Hospital Affiliated to Shanghai Jiao Tong University;Collaborative Innovation Center for Biotherapy,West China Hospital,Sichuan University;Key Laboratory of Embryo Molecular Biology,Ministry of Health & Shanghai Key Laboratory of Embryo and Reproduction Engineering;  
As a gene delivery system,Lentiviral vector( LVV) has been being increasingly used in gene therapy trials,and drawn large attention in recent years. Yet,transcriptional read-through( TRT) is one of potential risks in its application. For the purpose of improving the detection methods of TRT,and making the results intuitive and accurate,a codon optimized reporter gene,Lac Z,was placed downstream to the 3'- LTR of the proviral vector to simulate its state in chromosome. Upon the transfection followed with Lac Z staining,the cells with TRT phenomenon would be stained blue,and the TRT rate would therefore be visualized at the translational level. Correlating well with the outcomes of q RT-PCR,the established methodology could reflect the feature of TRT,which paves a way for improving LVV biosafety for clinical applications
【Fund】: 国家自然科学基金资助项目(81271690)
【CateGory Index】: R373;Q78
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1 Jiaping He1,Yudan Fang1,2,Fan Zhang1,Fengqiang Sun1,Juan Wang1,2,and Jingzhi Zhang1,2 1 Shanghai Institute of Medical Genetics,Children’s Hospital of Shanghai,Children’s Hospital Affiliated to Shanghai Jiao Tong University,Shanghai 200040,China 2 Key Laboratory of Embryo Molecular Biology,Ministry of Health & Shanghai Key Laboratory of Embryo and Reproduction Engineering,Shanghai 200040,China;Establishment of the methodology for quantifying lentiviral vector transcriptional read-through rate[J];Chinese Journal of Biotechnology;2013-07
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1 SUN Yong1,TANG Bi-kui1,2,BAI He-lu1,ZHU Nai-shuo1(1.Department of Microbiology and Microbial Engineering,School of Life Sciences,Fudan University, Shanghai 200433,China;2.Department of Life Sciences,Bengbu Medical College,Bengbu 233030,China);Expression of HBV-X Gene Induced by Lentiviral Vector in Mice[J];Journal of Fudan University(Natural Science);2013-04
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