RNA Interference (RNAi) technology has rapidly become one of the key methods used in functional genomics. RNAi is used to block the expression of genes and create phenotypes that can potentially yield clues about the function of these genes. In the postgenomic era, the elucidation of the physiological function of genes has become the rate-limiting step in the quest to develop 'gene-based drugs' and RNAi could potentially play a pivotal role in the validation of such novel drugs. In this overview, the basic concepts and applications of RNAi biology are discussed. Leading experts from both…mehr
RNA Interference (RNAi) technology has rapidly become one of the key methods used in functional genomics. RNAi is used to block the expression of genes and create phenotypes that can potentially yield clues about the function of these genes. In the postgenomic era, the elucidation of the physiological function of genes has become the rate-limiting step in the quest to develop 'gene-based drugs' and RNAi could potentially play a pivotal role in the validation of such novel drugs. In this overview, the basic concepts and applications of RNAi biology are discussed. Leading experts from both academia and industry have contributed to this invaluable reference. The volume is forwarded by Andrew Fire, one of the winners of the 2006 Nobel Prize for the discovery of RNA Interference.
Part I. Basic RNAi, siRNA and Gene Silencing Mechanisms: 1. RNAi beginnings: overview of the pathway in C. elegans 2. Dicer in RNAi: its roles in vivo and utility in vitro 3. Genes required for RNAi 4. MicroRNAs: a small contribution from worms 5. miRNAs in the brain and the application of RNAi to neurons Part II. Design and Synthesis of siRNAs: 6. Design and synthesis of small interfering RNA (siRNA) 7. Automated design and high throughput synthesis of siRNA 8. Rational design of siRNAs with the Sfold software 9. Enzymatic production of siRNAS Part III. Vector Development and In Vivo, In Vitro and In Ovo Delivery Methods: 10. Six methods of inducing RNAi in mammalian cells 11. Viral delivery of siRNA 12. siRNA delivery by lentiviral vectors: design and applications 13. Liposomal delivery of siRNAs in mice 14. Chemical modifications to achieve increased stability and sensitive detection of siRNA 15. RNAi in postimplantation mouse embryos 16. In ovo RNAi opens new possibilities for functional genomics in vertebrates Part IV. Gene Silencing in Model Organisms: 17. Practical applications of RNAi in C. elegans 18. Inducible RNAi as a forward genetic tool in Trypanosoma brucei 19. RNAi gene silencing in yeast 20. RNA silencing in filamentous fungi: Mucor circinelloides as a model organism 21. RNAi and gene silencing phenomenon mediated by viral suppressors in plants Part V. Drug Target Validation: 22. Delivering siRNA in vivo for functional genomics and novel therapeutics 23. The role of RNAi in drug target validation: application to Hepatitis C 24. RNAi in the drug discovery process 25. RNAi: use in the drug discovery process Part VI. Therapeutic and Drug Development: 26. RNAi-mediated silencing of viral gene expression and replication 27. RNAi in drug development: practical considerations 28. RNAi for the treatment of liver diseases 29. RNAi applications in animal systems Part VII. Genome Wide RNAi and High Through-Put Analysis: 30. High through-put RNAi-by-soaking in Caenorhabditis elegans 31. Tools for integrative genomics: genome-Wide RNAi and expression profiling in Drosophila 32. Microarray analysis and RNA silencing to determine genes functionally important in Mesothelioma 33. High through-put RNAi 34. Generation of highly specific vector-based shRNAi libraries directed against the entire human genome.
Part I. Basic RNAi, siRNA and Gene Silencing Mechanisms: 1. RNAi beginnings: overview of the pathway in C. elegans 2. Dicer in RNAi: its roles in vivo and utility in vitro 3. Genes required for RNAi 4. MicroRNAs: a small contribution from worms 5. miRNAs in the brain and the application of RNAi to neurons Part II. Design and Synthesis of siRNAs: 6. Design and synthesis of small interfering RNA (siRNA) 7. Automated design and high throughput synthesis of siRNA 8. Rational design of siRNAs with the Sfold software 9. Enzymatic production of siRNAS Part III. Vector Development and In Vivo, In Vitro and In Ovo Delivery Methods: 10. Six methods of inducing RNAi in mammalian cells 11. Viral delivery of siRNA 12. siRNA delivery by lentiviral vectors: design and applications 13. Liposomal delivery of siRNAs in mice 14. Chemical modifications to achieve increased stability and sensitive detection of siRNA 15. RNAi in postimplantation mouse embryos 16. In ovo RNAi opens new possibilities for functional genomics in vertebrates Part IV. Gene Silencing in Model Organisms: 17. Practical applications of RNAi in C. elegans 18. Inducible RNAi as a forward genetic tool in Trypanosoma brucei 19. RNAi gene silencing in yeast 20. RNA silencing in filamentous fungi: Mucor circinelloides as a model organism 21. RNAi and gene silencing phenomenon mediated by viral suppressors in plants Part V. Drug Target Validation: 22. Delivering siRNA in vivo for functional genomics and novel therapeutics 23. The role of RNAi in drug target validation: application to Hepatitis C 24. RNAi in the drug discovery process 25. RNAi: use in the drug discovery process Part VI. Therapeutic and Drug Development: 26. RNAi-mediated silencing of viral gene expression and replication 27. RNAi in drug development: practical considerations 28. RNAi for the treatment of liver diseases 29. RNAi applications in animal systems Part VII. Genome Wide RNAi and High Through-Put Analysis: 30. High through-put RNAi-by-soaking in Caenorhabditis elegans 31. Tools for integrative genomics: genome-Wide RNAi and expression profiling in Drosophila 32. Microarray analysis and RNA silencing to determine genes functionally important in Mesothelioma 33. High through-put RNAi 34. Generation of highly specific vector-based shRNAi libraries directed against the entire human genome.
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