RNA Interference Technology

Krishnarao Appasani is the Founder of GeneExpression Systems, a gene discovery company focusing on functional genomics in cancer research.

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.