BIT 471: 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:

  1. Introduction to RNAi
  2. Post Transcriptional Gene Silencing in Plants
  3. C.elegans: RNAi History and Experiments
  4. Drosophila: RNAi Background and Experiments
  5. RNAi and Mammalian Systems
  6. MicroRNAs, Antisense RNA and Other RNA Silencers
  7. Therapeutic Uses of RNA

Labs:

  1. Preparation of DNA-Directed Silencing Constructs 1 week
  2. Virus Induced Gene Silencing of Magnesium Chelstase, a Chlorophyll Biosynthesis Enzyme, in Tobacco Plants. Real Time RT-PCR Assay 5 weeks
  3. C. elegans RNAi Experiment to Knockdown Transgenic GFP Expression via Feeding of Silencing Constructs. Qualitative Phenotypic Assesment 2 weeks
  4. Transient Transfection Delivery of Short Hairpin RNA Expression Plasmids to Mammalian Cell Culture. Semi-quantitative Western Blot. 3 weeks

Back to Courses