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噬菌体phi29 pRNA载体在RNA干扰中的初步应用研究

Bacteriophage phi29 pRNA as Small Interference RNA Vector in RNA Interference
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摘要 Gene-silencing siRNA has shown great promise for the treatment of genetic diseases,cancers,and viral infections,but its therapeutic value has been hindered by the lack of an efficient and safe in vivo delivery system to target specific cells.The motor pRNA of phi29 has a strong tendency to form dimers,trimers and hexamers by interaction of interlocking right and left hand loops.This unique feature makes pRNA an ideal vector for the delivery of multiple therapeutic RNAs.Toxicity of pRNA was detected by transfection of 8 pRNA in HeLa cells.A pRNA-based vector was designed to carry siRNAs to inhibit GFP or HBV surface gene expression.Silence effects on siRNA against expression of GFP or HBV surface gene were detected in HeLa cells.Viral replicative intermediates were detected by Southern blotting.The results of toxicity study showed there was no toxicity of pRNA to cultured monolayer cells.The siRNA connected with pRNA can inhibit GFP or HBV surface gene expression in HeLa cells and inhibit HBV replication in HepG2 cells.These data suggest that pRNA can be used as a vector for imparting stability to siRNA in vitro. Gene-silencing siRNA has shown great promise for the treatment of genetic diseases, cancers, and viral infections, but its therapeutic value has been hindered by the lack of an efficient and safe in vivo delivery system to target specific cells. The motor pRNA of phi29 has a strong tendency to form dimers, trimers and hexamers by interaction of interlocking right and left hand loops. This unique feature makes pRNA an ideal vector for the delivery of multiple therapeutic RNAs. Toxicity of pRNA was detected by transfection of 8 pRNA in HeLa cells. A pRNA-based vector was designed to carry siRNAs to inhibit GFP or HBV surface gene expression. Silence effects on siRNA against expression of GFP or HBV surface gene were detected in HeLa cells. Viral replicative intermediates were detected by Southern blotting. The results of toxicity study showed there was no toxicity of pRNA to cultured monolayer cells. The siRNA connected with pRNA can inhibit GFP or HBV surface gene expression in HeLa cells and inhibit HBV replication in HepG2 cells. These data suggest that pRNA can be used as a vector for imparting stability to siRNA in vitro.
出处 《病毒学报》 CAS CSCD 北大核心 2007年第2期148-152,共5页 Chinese Journal of Virology
基金 国家自然科学基金资助项目(30571646) 国家教育部博士学科点科研基金(20030487070)
关键词 噬菌体 PRNA RNA干扰 乙型 肝炎病毒 bacteriophage vector RNA interference hepatitis B virus
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