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RNAi RNA interference

Rebeckah Ruiz. RNAi RNA interference. In this Presentation. What is RNAi Overview of RNAi History and Discovery of RNAi Applications of RNAi Future Implications. z. What is RNA Interference. A conserved biological response mechanism that inhibits gene expression prior to translation.

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RNAi RNA interference

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  1. Rebeckah Ruiz RNAiRNA interference

  2. In this Presentation • What is RNAi • Overview of RNAi • History and Discovery of RNAi • Applications of RNAi • Future Implications

  3. z What is RNA Interference • A conserved biological response mechanism that inhibits gene expression prior to translation. • Triggers of the RNAi pathway include foreign DNA or dsRNA of viral origin, transposons, and pre-microRNA. • Targets mRNA for a specific gene, stopping all production of a certain protein. • A safety mechanism developed to protect us. • Plays a role in regulating development

  4. z z But How Does RNAi Work? A case study in Petunias. • Rich Jorgensen at the University of Arizona in 1990 wanted to make a novel type of Petunia with a deep purple color. • His group isolated the “purple gene” and tried expressing extra copies of the same gene by introducing them back into the genome • What he got? Completely white flowers. • The very gene he wanted to overexpress was turned off completely? • Post Transcriptional Gene Silencing • This was the first clue to the RNAi pathway. • Similar Phenomena in Nuerospora; quelling • Took 10 years to truly understand what was happening.

  5. Craig Mello and Andrew Fire “A pair of Americans won the coveted Nobel Prize in medicine or physiology for 2006 today for their groundbreaking discovery that strands of RNA can selectively silence genes. The finding has revolutionized genetics, offered new insights into cellular behavior, and energized medical research. Craig Mello of the University of Massachusetts (U Mass) Medical School in Worcester, and Andrew Fire of Stanford University in California learned early this morning that the $1.4 million prize was theirs” Phenotypic effect after injection of single-stranded or double-stranded unc-22 RNA into the gonad of C. elegans. The unc-22 gene encodes a myofilament protein.

  6. “The biochemistry of RNAi was further elucidated in an in vitro system based on Drosophila embryo extracts” The Dicer enzyme recognizes dsRNA and chops it into short interfering RNAs (siRNAs) A single base pair difference between the siRNA template and the target mRNA is enough to block the process. The siRNAs are picked up by the RNA induced silencing complex (RISC) and attached The siRNA-RISC complex can now be used to find and attach to target mRNA (complimentary to siRNA) and chops the mRNA into small pieces The chopped up mRNA can no longer be used to create proteins and is not functional

  7. AKA The Cutest Biological Pathway Ever Double stranded RNA RISC SLICER protein

  8. z z Applications of RNAi • Hematology • Oncology • Stem cell biology • Infectious diseases • Agriculture • Gene identification • Gene Therapy • Many more

  9. RNA Interference: Hepatitis C • Anton McCaffrey and Colleagues at the University of Stanford in 2002 • Investigated whether RNAi could be directed against a human pathogenic RNA expressed in a mouse, hepatitis C virus (HCV). • Infection by HCV, an RNA virus that infects 1 in 40 people worldwide, is the most common reason for liver transplantation in the United States and Europe. HCV siRNA reduced expression by 75% (+- 6.8%; 6 animals per group). (p<0.0115). This result suggests that it may be feasible to use RNAi as a therapy against other important human pathogens

  10. Oncology Schultheis, Beate, et al. “First-in-Human Phase I Study of the Liposomal RNA Interference Therapeutic Atu027 in Patients With Advanced Solid Tumors.” Journal of Clinical Oncology, Dec. 2014 • Atu027 is a novel liposomal RNAi therapeutic to inhibit local tumor invasion • Thirty-four patients with advanced solid tumors (pulmonary metastasis) received 10 escalating doses of Atu027 as one single followed by eight intravenous infusions twice per week during a 28-day cycle. • Response was monitored by computed tomography/magnetic resonance imaging at baseline, at the end of treatment. • Disease stabilization was achieved in 41% of patients at end of treatment and some experienced complete or partial regression of metastases.

  11. Conclusion • RNAi is a powerful tool for analyzing, identifying, and silencing gene function • Leading treatments for cancer and other devastating diseases • Silence genes required for cell proliferation • Silence genes encoding key structural proteins • High specificity; cost effective • Easily labelled and introduced into patients • Simultaneously perform experiments in multiple cell types of interest • Viruses are basically pirates…

  12. References McCaffrey, Anton P., et al. “RNA Interference in Adult Mice.” Nature, vol. 418, no. 6893, July 2002, p. 38. EBSCOhost,libproxy.unm.edu/login?url=https://search.ebscohost.com/login.aspx?direct=true&db=a9h&AN=6982179&site=eds-live&scope=site. Schultheis, Beate, et al. “First-in-Human Phase I Study of the Liposomal RNA Interference Therapeutic Atu027 in Patients With Advanced Solid Tumors.” Journal of Clinical Oncology, vol. 32, no. 36, Dec. 2014, pp. 4141–4148. EBSCOhost, doi:10.1200/JCO.2013.55.0376. Scott Q. Harper, et al. “RNA Interference Improves Motor and Neuropathological Abnormalities in a Huntington’s Disease Mouse Model.” Proceedings of the National Academy of Sciences of the United States of America, no. 16, 2005, p. 5820.  Napoli C, Lemieux C, Jorgensen R. Introduction of a chimeric chalcone synthase gene into petunia results in reversible co-suppression of homologous genes in trans. The Plant Cell. 1990;2:279–85. Romano N, Macino G. Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences. MolMicrobiol. 1992;6:3343–53. Fire A, Xu S, Montgomery MK, Kostas SA, Driver SE, Mello CC. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 1998;391:806–11. Obbard DJ, Gordon KHJ, Buck AH, Jiggins FM. The evolution of RNAi as a defence against viruses and transposable elements. Philosophical Transactions of the Royal Society B: Biological Sciences. 2009;364:99–115. Antler, Christine. “Antisense RNA” August 2003. The Science Creative Quarterly RNAi: Slicing, dicing and serving your cells - Alex Dainis Ted-Ed Published on Aug 12, 2013

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