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  • 09월 04일 13시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

Suppression of HCV genome replication in hepatic cells with exogenous shRNA or shRNA expression system

2009년 8월 14일 15시 59분 26초
36P217포 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
목 <발표Ⅱ>
저자 및
김경보, 이복희, 김동은1
건국대학교 생명공학, Korea
1건국대학교 생명공학과, Korea
RNA interference (RNAi) is a highly conserved gene silencing mechanism among mammalia. This cellular process requires small duplex RNA to degrade target mRNA in sequence specific manner. At present, RNAi has been appled for functional genomics studies as well as potential therapeutic agent. Short hairpin RNAs (shRNA) are commonly used for RNAi. The shRNA hairpin structure is cleaved by the cellular machinery into siRNA, which is then bound to the RNA-induced silencing complex (RISC). Previous studies in our laboratory discovered DNAzymes that very efficiently cleave hepatitic C viral genome in hepatic cells. We suggested that target sites of the DNAzymes are accessible to antisense oligonucleotide (AON). In this study, we designed shRNA targeting against this accessible site and proved shRNA mediated suppression of hetatitic C viral replication in hepatic cells. We utilized two kinds of shRNAs to eradicate the target in the cells; lentiviral vector system and exogenous shRNA transcribed with T7 RNA polymerase. Recently, many studies have shown that innate immune response is triggered by these kinds of duplex small RNA. The immune response can cause inhibition of gene expression in a sequence-independent manner. Thus, we now have been examining whether our designed shRNA and shRNA expression vector induce the Toll like receptor (TLR) mediated immune response when we transfect the molecule into hepatic cell. Furthermore, we will compare the target silencing ability of these shRNA systems in hepatitis cells containing HCV replicon RNA.