SUBJECT

Transgenic Organisms: GMOs, Gene Therapy, Knockout, Live Imaging

lecture

master

Faculty of Science

Biology MSc

2

Semesters 1-4

Autumn/Spring semester

1. week - Introduction: definitions, types and uses of transgenic systems.Basic methods in transgenesis, such as recombinant DNA techniques.

2. week - Genome projects and benefits. Ethical issues. Historical overview.

3. week - Microbial biotechnology. The living cell as a bioreactor. PGPR bacteria, mycorrhiza projects, bioinsecticides. Plant biotechnology. Generation of GM plants to enhance biotic/abiotic stress tolerance, modify development and metabolism (terminator technology, Golden Rice, edible vaccine).

4. week - Animal biotechnology. Generation of transgenic animals (microinjections, GM ES cell chimaeres, cloning). Major applications: production of therapeutic proteins in milk, xenotransplantation.

5. week - Transgenic methods in therapy. Diagnostic applications, recombinant drugs, gene therapy, GM stem cell therapy.

6. week - Transgenic methods in research. Mutant rescue, ecopic and overexpression, transpozon mutagenesis (eg. enhancer trapping). The uses of fusion proteins such as GFP.

7. week - Transgenic methods in yeast and worm research. Genome-wide knockout, knockdown, and GFP-fusion libraries. Colocalisation projects.

8. week - Transgenic methods in Drosophila research. Uses of the P element. The Gal4-UAS system. Somatic mutant and overexpression clones in mosaic analysis. Gene knockout and transgenic RNAi.

9. week - Cell culture. Transient and stable transfections, fusion proteins, dominant-negative and siRNA studies. Homologous recombination-based gene targeting (insertion or replacement knock-out, knock-in) in mouse embryonic stem cells.

10. week - KO and transgenic mouse models. Temporal/spatial control of gene expression and deletion (tet-on and tet-off systems, Cre-lox techniques). Genetic models of physiology and disease ("cells don't have blood pressure").

11. week - Transgenic and KO models of monogenic and complex human diseases, such as Lesch-Nyhan syndrome, cystic fibrosis, neurodegeneration and cardiovascular diseases, tumor formation etc.

12. week - Microscopy techniques. Imaging fluorescent molecules in live cells, time-lapse videos, FRAP, FLAP, FLIP, FRET, photoactivation.

13. week - Superresolution fluorescent microscopy. STED, STORM, 3D-SIM etc. systems.

14. week - Final written test.

• J. Tomiuk, K. Wöhrmann, Andreas Sentker: Transgenic Organisms, Birkhäuser, 2012

• Carl A. Pinkert: Transgenic Animal Technology, Newnes, 2014