Construction of Recombinant Adenovirus with siRNA Against Collagen I

doi:10.3969/j.issn.1000-565X.2013.12.016

Journal of South China University of Technology (Natural Science Edition) ›› 2013, Vol. 41 ›› Issue (12): 95-100.doi: 10.3969/j.issn.1000-565X.2013.12.016

• Biological Engineering • Previous Articles Next Articles

Construction of Recombinant Adenovirus with siRNA Against Collagen I

Yao Yong- chang Zhou Jia- an Chen Xiao- feng

School of Materials Science and Engineering,South China University of Technology,Guangzhou 510640,Guangdong,China∥National Engineering Research Center for Tissue Restoration and Reconstruction,Guangzhou 510006,Guangdong,China

Received:2012-10-23 Revised:2012-12-13 Online:2013-12-25 Published:2013-11-19
Contact: 陈晓峰(1956-)，男，博士，教授，主要从事生物医学材料研究． E-mail:chenxf@scut.edu.cn
About author:姚咏嫦(1979-)，女，博士，讲师，主要从事基因传递在组织再生的应用研究．E-mail:yaoyc@scut．edu．cn
Supported by:
国家自然科学基金青年科学基金项目(51303058);华南理工大学中央高校基本科研业务费专项资金资助项目(2011ZM0010);广东省自然科学基金资助项目(S2011040000208);教育部留学回国人员科研启动基金资助项目

Abstract

Abstract:

As compared with the stem cells from other sources,synovium- derived mesenchymal stem cells(SMSCs) are of higher proliferative and chondrogenic ability.However,SMSCs express Collagen I inherently,which may impair the overall quality of the regenerated counterpart.In order to solve this problem,first,shRNAagainst Collagen I was designed and constructed in the adenoviral vector.Next,linear recombinant adenoviral vectorwas generated by the digestion with enzyme PacI and was used to transfect 293 cells for package. Then,the titrationof recombinant adenovirus was determined,and the corresponding amplification and purification were performed toobtain recombinant adenovirus with high titration.Moreover,fibroblasts and osteoblasts were respectively infectedby the recombinant adenovirus,and fluorescence microscopy was adopted to detect the transfection efficiency.Fi-nally,real- time quantitative PCR experiments were carried out to verify the targeting effect of siRNA.Experimentalresults show that the packaged recombinant adenoviruses are of high transfection efficiency and can successfully en-code siRNA against Collagen I.

Key words: synovium- derived mesenchymal stem cell, adenovirus, siRNA, Collagen I, vector, transfection

CLC Number:

Q782

Yao Yong- chang Zhou Jia- an Chen Xiao- feng. Construction of Recombinant Adenovirus with siRNA Against Collagen I[J]. Journal of South China University of Technology (Natural Science Edition), 2013, 41(12): 95-100.

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Construction of Recombinant Adenovirus with siRNA Against Collagen I

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