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Combinatorial method to screen RNA-cleaving oligodeoxyribozyme and inhibitory RNA aptamer for therapeutic application

2009년 8월 14일 11시 21분 14초
금15H3심 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
금 10시 : 10분
생명화학 - Nanomaterials and Nucleic Acids for Molecular Therapy
저자 및
김동은, 박꽃한아름
건국대학교 생명공학과, Korea
Nucleic acids provide an enormous potential as a therapeutic reagent against viral diseases and cancers. Especially, RNA-cleaving oligodeoxyribozyme and RNA aptamer against pathogenic proteins are promising tools as therapeutic nucleic acids. A class of antisense oligodeoxyribozymes, known as the 10-23 DNAzyme, has been shown to efficiently cleave target RNA at purine-pyrimidine junctions in vitro. A screening strategy was performed to identify accessible cleavage sites for DNAzyme in the target RNA, Hepatitis C virus nonstructural gene 3 (HCV NS3) that encodes viral helicase and protease, starting from a pool of random DNAzyme library. The screened DNAzymes, when transfected to the human hepatoma cells harboring the HCV subgenomic replicon RNA, efficiently inhibited HCV RNA replication in cells by reducing expression of HCV NS3 RNA and protein. Thus, the selected oligonucleotides as well as the selection strategy can be applicable for a new class of anti-HCV drugs as antisense oligonucleotides-based therapy. The same approach with the DNAzyme screening has been executed to inhibit beta-catenin expression in the colon cancer cells. The selected DNAzymes have shown effective degradation of beta-catenin RNA and subsequent expression of the gene in colon cancer cells. Another example of oligonucleotide-based molecular therapy is RNA aptamers that specifically bind and inhibit the activity of certain pathogenic protein. RNA aptamers have been isolated against SARS coronavirus (SCV) NTPase/Helicase from RNA library containing random sequences of 40 nt using in vitro selection technique (SELEX). The isolated RNAs were observed to efficiently inhibit double-stranded DNA unwinding activity of the helicase. These results suggest that the pool of selected aptamers might be potentially useful as anti-SCV reagents.