e-Day Lesson #3 Advanced Genetics and Biotechnology Investigation of RNAi (Interference RNAs) Name_____________ Part A.

Watch Nova Science NOW: 8-RNAi at http://www.youtube.com/watch?v=Vh3NHdjnyQ or http://www.pbs.org/wgbh/nova/body/rnai.html and then answer these questions: 1) What do each of these represent in the video clip? What is the role of each? a) monks RNA. Transcribes the DNA and sends it out to be used outside the nucleus. b) cooks Ribosomes. They take the genetic info and build proteins with them. c) cops RNAi. They locate and kill viruses, stopping them from replicating. d) pirate ship Viruses. They invade the cells, forcing them to create copies of themselves. 2) Why did the petunias turn white when scientists were trying to make a darker purple petunia? The genes added to increase the amount of purple was viewed as an offensive virus by the RNAi. They destroyed these genes, then recognised other pigment genes as similar, hence killing those too. With all pigment genes destroyed, the flowers no longer made purple pigments. 3) Why do cells naturally have RNAi s? To protect against viruses. The RNAi defend against them, recognising viruses and similar shapes, destroying them. 4) How are RNAi being used to help the woman with macular degeneration? She had genes inserted into her cells that coded for additional blood vessels. The RNAi recognised these genes as virus genes, killing them and other, similar genes, hence reducing the amount of blood vessels made. 5) How do scientists think that RNAi s will be useful in the future? Will have the capability to target any disease in the future. Also will give the ability to turn off and discover the use of any gene, resulting in genetic engineering.

Part B: Most of the scientific articles about current RNAi research and technology are highly technical. Click on the following link to access the article Current Prospects for RNA-Interference Based Therapies and locate Table 1 and Table 2 within the article. http://www.societyns.org/runn/2013/pdfs/DAVIDSONBLD%20and%20McCray%20NRG %202011.pdf

1) Using Table 1 Clinical Trials for RNAi Therapy: a) List the five clinical settings for current clinical trials. Ocular and retinal disorders, cancer, kidney disorders, LDL lowering, antiviral b) Choose three of the disorders listed in the indicator column and using other sources, define what each disorder/disease is. Choose indicators that are not familiar to you. This will help you understand the types of clinical situations that scientists hope RNAi will be able to treat. Glaucoma- a group of ocular disorders that can cause permanent eye damage. Increased fluid pressure in the eyes, resulting in optic nerve damage. Hypercholesterolemia- presence of high cholesterol levels in the blood. Metastatic melanoma- when melanoma has spread to distant regions, affecting many organs. Melanoma is the forming of melanin-forming tumor cells. 2) Look at Table 2 Methods for the Delivery of RNAi Triggers to Cells and Tissues. This table lists the various ways that scientists are using to insert the RNAi components into cells. Notice under the Viral Vectors packaging capacity is listed. This corresponds to what we discussed in Section 14.1 of your textbook. After reviewing Table 2, answer the following questions: a) Which of the viruses has the largest packaging capacity? What is this virus normally associated with? Herpes simplex virus. HSV-1 produces cold sores, whereas HSV-2 is an STD. b) Escherichia coli and Salmonella typhymurium deliver the siRNA to what type of tissue? What are these bacteria normally associated with? Gut tissue. Both are seen as potentially dangerous viruses, associated with food poisoning.

c) Cholesterol and nanoparticles are listed under Non-viral Formulations. What advantages do you think would there be to using a non-viral or bacterial vector? More stable, ability to deliver to multiple types of cells.