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Overview:
- 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
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