Journal of South China University of Technology(Natural Science Edition) ›› 2012, Vol. 40 ›› Issue (1): 114-119.

• Biological Engineering • Previous Articles     Next Articles

Package and Verification of Adeno-Associated Virus Vector Constructed with siRNA Targeting TTF-1

Gao Qiang  Cao Yi-cheng  Yang Lei   

  1. School of Biological Science and Engineering,South China University of Technology,Guangzhou 510006,Guangdong,China
  • Received:2011-07-07 Revised:2011-10-11 Online:2012-01-25 Published:2011-12-01
  • Contact: 曹以诚(1949-) ，男，教授，博士生导师，主要从事生物医药开发和生物信息学研究．E-mail: yccao@scut.edu.cn E-mail:gaoqiang201@yahoo.com.cn
  • About author:高强(1973-) ，男，博士生，讲师，主要从事生物医药开发方面的研究．
  • Supported by:

    国家自然科学基金重大研究项目( 90412015) ; 教育部中国网格计划生物信息网格平台子项目( B12137040130) ;广州市科技攻关计划项目( 2005Z12E4023)

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Abstract:

In order to introduce siRNA targeting TTF-1 gene into lung adenocarcinoma cells,first,three TTF-1-gene-specific siRNAs for NCI-H1975 cells were respectively designed and constructed in adeno-assosiated virus( AAV) . Next,the primary culture liquid of AAV was harvested after the package of AAV in 293 cells. Then,thetiter of AAV was detected after the concentration by ultrafiltration and after the purification by column chromatography for the primary culture liquid of AAV. Finally,NCI-H1975 cells were infected by recombinant AAV,nd the interference effect of siRNA and the cell biology effect were respectively verified by Western-Blot experiments and cells apoptosis experiments,ith good interference effect of siRNA and apoptosis of NCI-H1975 being both revealed,which means that infective AAV-TTF-1-siRNA was successfully constructed.

Key words: TTF-1, deno-associated virus, mall interfering RNA, ector, nfection verification

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