Short interfering RNA siRNA

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Presentation transcript:

Short interfering RNA siRNA 12/5/2018

Part I: Gene silencing exist in many multicellular organisms fruitfly Drosophila roundworm C. elegans fungus Neurospora 12/5/2018

Gene silencing go back to the earliest epochs of life on Earth Wang, Kumar & Hedges, 1999 This ancient immune system seems conserved over 1.5 billion years ...thus we can learn about different aspects of gene and RNA silencing wherever it is easiest... plants... roundworms... flies... human cell lines... 12/5/2018

A brief history of RNA silencing 1998: Andrew Fire and colleagues show that double-stranded RNA induce gene silencing in C. elegans 1999: Hamilton & Baulcombe find small RNA where genes are silenced in plants 2000: Greg Hannon and colleagues isolate Dicer, the siRNA producing RNase-III enzyme in Drosophila 2001: Thomas Tuschl and colleagues show that synthetic small RNA introduced into human cells can silence human genes 12/5/2018

13.10 RNA Interference Is Related to Gene Silencing RNA interference triggers degradation of mRNAs complementary to either strand of a short dsRNA. 12/5/2018 Figure 13.21

Model for RNAi mechanism Dicer (RNA-induced silencing complex) Initiation step Effector step 12/5/2018 Hammond et al., 2001

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Produzione di siRNA Brummelkamp, 2002; Paddision, 2002; Miyahishi, 2002; Paul, 2002; Sui, 2002; Lee, 2002; Jacque, 2002; Yu, 2002; Yang, 2002 12/5/2018

Inhibition of cholesterol synthesis by siRNA: A recent study by Alnylam Inc "...Administration of chemically modified siRNAs resulted in silencing of the apoB messenger RNA in liver and jejunum, decreased plasma levels of apoB protein, and reduced total cholesterol. " Soutschek et al, Nature, 2004 12/5/2018

More small RNA exist stRNA (short temporal RNA, also called micro-RNA, miRNA) inhibit mRNA translation (Lee & Ambros, 1993; Reinhart, 2000; Lee, 2001; Lau, 2001; Lagos-Quintana, 2001; Llave, 2002; Lim, 2003) microRNA comes from microGenes, transcripted as ~70 nt hairpins, and processed (as is shRNA) to short ~21-mer RNAs... 12/5/2018

13.9 MicroRNAs Are Regulators in Many Eukaryotes Animal and plant genomes code for many short (~22 base) RNA molecules called microRNAs. MicroRNAs regulate gene expression by base pairing with complementary sequences in target mRNAs. 12/5/2018

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