Module 2 Expression Engineering 20 109 Lecture 3 October 18th 2007 Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY STEM Education best way to achieve Adequate Yearly Progress Content removed due to copyright restrictions Excerpt from Medina Jennifer Making Cash a Prize for High Scores on Advanced Placement Tests The New York Times October 15 2007 http www nytimes com 2007 10 15 nyregion 15rewards html accessed May 8 2008 Excerpt from Kula Adam and Alexandra Frean Make science easier examiners are told The Times August 29 2007 http www timesonline co uk tol news uk education article2344054 ece accessed May 8 2008 Excerpt from Cavanaugh Sean Federal Rule Yields Hope for Science Education Week published online October 5 2007 Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Expression Engineering Experiment Day 1 Day 2 Day 3 Image of glowing luciferase in a microcentrifuge tube removed due to copyright restrictions RT Figure by MIT OpenCourseWare Day 6 Day 5 Day 4 Image credits Day 1 Courtesy of Arkitek Studios Used with permission Day 2 Courtesy of The Exploratorium The Exploratorium http www exploratorium edu Day 4 Figure by MIT OpenCourseWare Day 5 Dr Natalie Kuldell Day 6 Courtesy of NIH Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Expression Engineering Experiment Lecture 1 Lecture 2 intro to cell culture transfection intro to gene exp n RNAi luciferase Lecture 3 Lecture 4 off target nonspecific RNAi Writing lecture Neal Lerner Lecture 5 Lecture 6 measuring gene express n microarray analysis Rebecca Fry Lecture 7 Lecture 8 high throughput technologies or review of your data RNAi applications no lab Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY RNAi Operation or Battleship Images removed due to copyright restrictions Photos of the games Operation and Battleship Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Where were you in 1998 Content removed due to copyright restrictions News related images France wins the World Cup Clinton under Investigation re Monika Lewinski President Clinton speaking at MIT commencement Movie poster for Titanic Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Source NIH Image removed due to copyright restrictions Excerpt from Table 1 Effects of sense antisense and mixed RNAs on progeny of injected animals In Fire A et al Potent and specific genetic interference by double stranded RNA in Caenorhabditis elegans Nature 391 19 February 1998 doi 10 1038 35888 Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY small RNA revolution of publications Image removed due to copyright restrictions Cover of Science 298 no 5602 20 December 2002 New roles for RNAs 15 in 1998 1000 in 2000 4500 in 2007 so far Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY RNA interference vocabulary Transfection siRNA Dicer RISC Courtesy of Arkitek Studios Used with permission Source RNAi Interface animation for nature com http www nature com focus rnai animations index html Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Nomenclature for siRNAs 1 TT 3 OH 5 PO4 3 OH TT 5 PO4 19 RNA AUG DNA ATG TAC UAA TAA ATT sense template coding strand template strand Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Nomenclature for siRNAs TT sense antisense TT RNA AUG DNA ATG TAC RISC UAA TAA ATT Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Nomenclature for siRNAs TT TT RNA AUG DNA ATG TAC RISC UAA TAA ATT Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY Delivery vs expression of dsRNA Methods of delivering double stranded RNA Organism Method C elegans Drosophila Trypanosomes dsRNA C elegans Drosophila Trypanosomes Plants Stem loop expression 5 3 dsRNA Trypanosomes Dual promoter 5 3 Plants 3 dsRNA 5 Virus Figure by MIT OpenCourseWare See Post transcriptional gene silencing by dsRNA Nature Reviews Genetics 2001 2 110 119 Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY RNAi siRNA vs miRNA RISC biochemistry unites RNAi Figure removed due to copyright restrictions fig from Svoboda review in Curr Opin Mol Ther 2007 9 3 248 Cite as Natalie Kuldell Course materials for 20 109 Laboratory Fundamentals in Biological Engineering Fall 2007 MIT OpenCourseWare http ocw mit edu Massachusetts Institute of Technology Downloaded on DD Month YYYY RNAi on paper Highly effective 90 reduction in gene expression Highly specific only targeted mRNA gets degraded Reproducible effects same with each expt l