miRNAs and siRNAs 5 th March 2013 Saeideh Jafarinejad 4/22/2013 Rhumatology Research Center Lab(RRC lab)

Non-coding versus Coding regions 4/22/2013Rhumatology Research Center Lab(RRC lab)

Non-coding RNAs A non-coding RNA (ncRNA) is a functional RNA molecule that is not translated into a protein.RNAtranslatedprotein Non-coding RNA Long noncoding RNA Small non-coding RNA microRNA piRNA Endo-siRNA rRNA tRNA snRNA 7SL RNA Y RNA TERC (telomerase RNA component) 4/22/2013Rhumatology Research Center Lab(RRC lab)

Small noncoding RNAs (small silencing RNAs) microRNA siRNA piRNA 4/22/2013Rhumatology Research Center Lab(RRC lab)

MicroRNAs Victor Ambros 4/22/2013Rhumatology Research Center Lab(RRC lab)

4/22/2013Rhumatology Research Center Lab(RRC lab)

microRNA, miRNA hsa-miR-21-a hsa-miR-12-1 hsa-mir-21 dme-miR-21 hpv-miR-21 5’Gppp5’G 3’AAAAA…A 4/22/2013Rhumatology Research Center Lab(RRC lab)

microRNAs Regulation of 50%of the mammalian protein-coding genes Spread throughout the human genome except the Y Chr.miRBase (19): 1600 precursors, 2042 mature 4/22/2013Rhumatology Research Center Lab(RRC lab)

The structure of human pri-miRNA 4/22/2013Rhumatology Research Center Lab(RRC lab)

microRNAs has-miR-21-5p hsa-mir-21-3p 4/22/2013Rhumatology Research Center Lab(RRC lab)

Mechanisms of target regulation by miRNAs 4/22/2013Rhumatology Research Center Lab(RRC lab)

Drosha Dicer Argonate Microprocessor RLC RNA Loading Co. RISC RNA Induced Silencing Co. 4/22/2013Rhumatology Research Center Lab(RRC lab)

Non-canonical pathways for microRNA production Drosha-independent pathways Mirtron miRNAs derived from snoRNAs miRNAs derived from endogenous short-hairpin RNAs (endo-shRNAs) and tRNA precursors tRNase Z-dependent pathways miRNAs derived from a terminal hairpin of an endo-siRNA precursor Dicer-independent pathways 4/22/2013Rhumatology Research Center Lab(RRC lab)

Mirtron 4/22/2013Rhumatology Research Center Lab(RRC lab)

4/22/2013Rhumatology Research Center Lab(RRC lab)

4/22/2013Rhumatology Research Center Lab(RRC lab)

RNAi (RNA interference) siRNA (small interferindg RNA) 4/22/2013Rhumatology Research Center Lab(RRC lab)

Endo-siRNA Plants & worms RdRP(+) Mammales & Flies RdRP(-) 4/22/2013Rhumatology Research Center Lab(RRC lab)

4/22/2013Rhumatology Research Center Lab(RRC lab)

Endo-siRNAs in flies and mammals 4/22/2013Rhumatology Research Center Lab(RRC lab)

Small Interfering RNAs siRNA piRNA siRNA (plants & fungi) miRNA piRNA Post transcriptional gene silencing (PTGS) Transcriptional gene silencing (TGS) 4/22/2013Rhumatology Research Center Lab(RRC lab)

4/22/2013Rhumatology Research Center Lab(RRC lab)

miRNAsiRNA Viruses,land plants, metazoan animals (+); fungi & choanofllagellates(-) All eukaryots Size: PTGSTGS,PTGS firstTranslation inhibition, later mRNA cleavage RNA cleavage Noncomplete base pairsComplete base pair endogenousEndo and Exogenous AGO protein relatedDicer and AGO related Is processed from stem-loop precursor with incomplete double- stranded character Are excised from long fully complementary double-stranded RNAs 4/22/2013Rhumatology Research Center Lab(RRC lab)

microRNA up-regulation Mimics synthetic short double-stranded oligonucleotides Expression Vectors 4/22/2013Rhumatology Research Center Lab(RRC lab)

Anti-miRNAs Synthetic, single strand, oligoneucleotides (AMO) Chemically modified cholesterol-conjugated single-strand oligos,or highdensity lipoprotein Vector conjugated inhinbition Sponges Eraser Mask 4/22/2013Rhumatology Research Center Lab(RRC lab)

Approaches for microRNA modulation and examples of application 4/22/2013Rhumatology Research Center Lab(RRC lab)

MicroRNAs Can Up-Regulate: 1) Switching from Repression to Activation microRNAs: Let-7 & miR369-3 mRNA: TNFα Site of function: AU-rich elements (AREs) & microRNA target site in 3′UTRs Condition: in cell cycle arrest 2) Mammalian miRNA target sites rarely occur within 5’UTRs. miR-10a induces, rather than inhibits, protein expression through binding to 5’UTRs of ribosomal protein transcripts. 3) miR-122 (the liver specific miRNA) binds to the 5′ UTR of Hepatitis C (HCV) genome and induces virus replication 4/22/2013Rhumatology Research Center Lab(RRC lab)

MicroRNAs in systemic rheumatic diseases 4/22/2013Rhumatology Research Center Lab(RRC lab)

4/22/2013Rhumatology Research Center Lab(RRC lab)

سپاس از توجه شما 4/22/2013Rhumatology Research Center Lab(RRC lab)

piRNA 4/22/2013Rhumatology Research Center Lab(RRC lab)