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RNA interference 1 RNA interference – Dicing Dicer Double-stranded RNA siRNAs (21-23 base pairs) RISC Helicase Slicer RNase RISC* (activated) mRNA siRNA: small interfering RNA RISC: RNAi silencing complex RISC*: activated RISC Slicer: it cleaves mRNA RNase: ribonuclease 2 RNA interference – amplification Double-stranded RNA Dicer siRNAs Dicer RNA-dependent RNA polymerase mRNA Amplified doublestranded RNA many siRNAs siRNA: small interfering RNA RISC: RNAi silencing complex RISC*: activated RISC Slicer: it cleaves mRNA RNase: ribonuclease 3 RNA interference DICER Ago + other RISC proteins activated RISC Ago + other RISC proteins Activated RITS complex DNA histone and DNA methylation DegradedRNA transcription repression RNA interference Andrew Z Fire; Craig C Mello C. elegans 1998 mex-3 gene Non-stained gfp gene non-injected (as a transgene in E. coli) Nutrition: double-stranded GFP mRNA expressing E. coli asRNA dsRNA Wild type RNAi-defected strain RNAi: degrading effect of double-stranded RNA on RNAs containing homologous sequences 4 RNA interference Andrew Z Fire; Craig C 5 Mello Technologies for selective down-regulation of genes Earlier: - antisense DNA oligonucleotide technology - Knock-out technology -----Today: knock down technology (RNA interference) Natural function: - antiviral mechanism - regulation of gene expression (?)
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