Dogmatic View of Gene Expression.pptVIP

Post-transcriptional Control: Non Coding RNAs: ‘RiboRegulators’ (~97% of RNAs Present in Human Cells are Non-Coding) RNA-Mediated Gene Silencing Discovery of RNAi Remarkable Properties of RNAi RNAi in Mammalian Cells MicroRNAs: Expanding Family of ‘RiboRegulators’ * Dogmatic View of Gene Expression DNA Protein RNA Quantitative Control: Levels of mRNA not proportional to levels of mRNA synthesized or protein produced Qualitative Control: More than one protein from a single gene (e.g. Differential RNA Processing or RNA editing) An “RNA-Centric” View of Gene Expression RNA DNA Protein RNA: A Diverse Class of Molecules RNA DNA rRNA tRNA snRNAs snoRNAs Guide RNA Catalytic: Ribozymes Telomerase Introns 5’ UTR 3’ UTR Vault Y RNAs 7SK Xist, H19 MicroRNAs Viral RNAs Retrotransposons Non Coding RNAs: SnoRNAs Large Family Intron-encoded Guide RNA Modification Non Coding RNAs: Telomerase RNA Component of telomerase Provides template for telomere synthesis Role in Cancer and Aging Post-transcriptional Gene Silencing (PTGS) or RNA Interference (RNAi)Transcriptional Gene Silencing (TGS)(RNA-dependent DNA Methylation)Gene Silencing By MicroRNAs Double-stranded RNA inject C. elegans Neg. control Uninjected Antisense RNA dsRNA sense antisense Nature 1998 391:806-811 Mex-3 mRNA detection in embryos by in situ hybridization dsRNA (not ssRNA) is interfering agent Sequence-specific loss of mRNA and protein Effective against exons not introns Potent (few dsRNA molecules/cell effective) Persistent (affects next generation) Effects can cross cell barriers (feed, soak) Mechanism of RNAi: Gene Silencing directed by ~22nt RNAs dsRNA ~22nt siRNAs target mRNA secondary siRNAs amplification processing degradation recognition copying + processing spreading Gene Silencing Factors C. elegans Drosophila Arabidopsis amplification processing degradation recognition copying + processing spreading DICER DCR-1 CAF AGO2 RDE-1 AGO1 RI