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BIT 471/571: RNA Interference

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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