Construction and identification of the recombinant adenovirus vector carrying a small interfering RNA targeting the peroxisome proliferator-activated receptor-γ

作     者：LIU Ming WANG Yi-sheng LI Yue-bai ZHAO Guo-qiang 

作者机构：Department of Orthopedic Surgery First Affiliated Hospital of Zhengzhou University Zhengzhou Henan 450052 China Basic Medical College Zhengzhou University Zhengzhou Henan 450001 China 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2012年第125卷第4期

页      面：671-675页

核心收录：

学科分类：0710[理学-生物学] 1002[医学-临床医学] 07[理学] 08[工学] 09[农学] 071007[理学-遗传学] 100214[医学-肿瘤学] 0901[农学-作物学] 0836[工学-生物工程] 090102[农学-作物遗传育种] 10[医学] 

主　　题：RNA interference peroxisome proliferator-activated receptor-γ adenovirus vector clone 

摘      要：Background Steroid-induced osteonecrosis of the femoral head （ONFH） is a common clinical disease,with a high disability *** present,efficient prevention and treatment of steroid-induced ONFH is still *** peroxisome proliferator-activated receptor-γ （PPARγ） is recognized as an important pathogenic gene for the development of steroid-induced *** interference （RNAi） is a tool for functional gene analysis,which has been successfully used to down-regulate the levels of specific target ***,down-regulation of PPARγ expression by RNAi may prevent the incidence of steroid-induced *** According to the principles of siRNA design,three duplex siRNA sequences （971-989,1253-1271 and 1367-1385） derived from the PPARy gene （NM_001082148） were *** duplexes were annealed,purified and ligated into 1.0-cytomegalovirus （CMV） shuttle *** shuttle vector was transfected into HEK293 *** HEK293 generated recombinant adenovirus vector carrying PPARγ siRNA sequences was purified and the titer of recombinant adenovirus was *** After the annealing of single-strand DNA oligo encoding short hairpin RNA （shRNA） sequences,products were identified by gel *** products were ligated into the 1.0-CMV shuttle vector and the recombinant shuttle vectors 1.0-CMV-971,1.0-CMV-1253 and 1.0-CMV-1367 were *** sequences of these recombinant vectors were *** then successfully constructed the recombinant adenovirus vector carrying siRNA targeting PPARγ.After purification,the virus titer was higher than 1010 plaque forming unit （PFU）/*** In this study,three recombinant adenovirus shuttle vectors carrying siRNA targeting PPARγ,including shuttle vectors 1.0-CMV-971,1.0-CMV-1253 and 1.0-CMV-1367,were successfully constructed and high titers of recombinant adenovirus were obtained.