Short
Courses | Day
1 | Day 2 | Download
Brochure
Tuesday, June 10
7:30am – 6:00 pm Registration Open
7:30 am Breakfast Workshop
(Sponsorships Available)
8:15 Chairperson’s Remarks
Richard Klinghoffer, Ph.D., Research Fellow, Biology, Rosetta Inpharmatics/Merck & Co., Inc.
8:20 Lentivirus-Mediated RNAi Screens
Richard Klinghoffer, Ph.D., Research Fellow, Biology, Rosetta Inpharmatics/Merck & Co., Inc.
To expand our capacity to interrogate disease-related gene families and signaling cascades by RNAi screens, we have established methods to produce and screen large collections of shRNA-containing lentiviral vectors. We have now used our methods to efficiently generate a screening compatible library of ~14,000 individually arrayed vectors targeting the human druggable genome (approximately 5,000 genes). Screens designed to identify novel regulators of disease have been performed and will be presented.
8:50 Rapid Creation of RNAi Libraries Using shRNA Microarrays
Michael C. Bassik, Ph.D., Post-doctoral Fellow, Laboratory of Michael T. McManus,
Department of Cellular and Molecular Pharmacology and Diabetes Center, Department
of Microbiology and Immunology, University of California, San Francisco
RNAi has become a widely used and powerful tool for conducting genetic screens. Lentiviral
shRNA delivery is most effective for stable gene disruption in diverse cell types, but currently available libraries are limited in a number of critical ways. To address some of these problems, we have developed a quick and efficient strategy to use microarray synthesis of complex shRNA pools in order to generate highly adaptable RNAi libraries. These libraries are inexpensive to create and maintain, can be easily changed to accommodate new vector design, and are less costly and cumbersome to use than standard arrayed libraries. Most importantly, this new strategy allows for much more comprehensive and effective gene targeting. We show here that we can couple the use of these libraries with sorting flow cytometry to identify and quantitate the potency of shRNAs against a number of targets, providing a significant improvement over current screening methods.
9:20 Rational Design Leads to More Potent RNA Interference Targeting
Hepatitis B Virus
Anton P. McCaffrey, Ph.D., Assistant Professor, Department of Internal Medicine, University of Iowa
Previously, we conducted proof-of-principle experiments using RNAi to degrade HBV RNAs in mice, and reduce levels of viral proteins and replicated DNA genomes. Recently we expressed the short hairpin RNA (shRNA), HBVU6#2, in HBV transgenic mice. While this RNAi trigger resulted in substantial HBV knockdown in mice, it also resulted in acute toxicity due to competition with the endogenous microRNA machinery. Using rational approaches based on mechanistic insights, we have designed RNAi triggers that are much more potent than the trigger identified in our original screen.
9:50 Networking Coffee Break, Poster and Exhibit Viewing
