Introduce students to the history of RNA Interference (RNAi) and its applications in common model organisms with a focus on the experimental design.
Students perform RNAi experiments in Nicotiana benthamiana (tobaccoa plants), in Caenorhabditis elegans (C. elegans) and in mammalian cell culture (human embryonic kidney cell line, HEK293).
Lectures:
Introduction to RNAi
Post Transcriptional Gene Silencing in Plants
C.elegans: RNAi History and Experiments
Drosophila: RNAi Background and Experiments
RNAi and Mammalian Systems
MicroRNAs, Antisense RNA and Other RNA Silencers
Therapeutic Uses of RNA
Labs:
Preparation of DNA-Directed Silencing Constructs -1 week
Virus Induced Gene Silencing of Magnesium Chelstase, a Chlorophyll Biosynthesis Enzyme, in Tobacco Plants. Real Time RT-PCR Assay -5 weeks
C. elegans RNAi Experiment to Knockdown Transgenic GFP Expression via Feeding of Silencing Constructs. Qualitative Phenotypic Assesment -2 weeks
Transient Transfection Delivery of Short Hairpin RNA Expression Plasmids to Mammalian Cell Culture. Semi-quantitative Western Blot. -3 weeks
