27541 - Advanced Molecular Genetics

Learning outcomes

The aim of this intensive course is to introduce students to molecular methodologies underlying functional genetics. Participants will be able to understand and plan experimental strategies to analize gene function through the use of transgenic animals.

Course contents

Classical and reverse genetics. Drosophila as a model organism. Balancer chromosomes as tools of Drosophila classical genetics. Phenotypic markers associated with balancer chromosomes.
EMS mutagenesis. Target-selected mutant screen by TILLING.
Transposable P elements  and hybrid dysgenesis. P Element-mediated germ-line transformation of Drosophila. Insertional mutagenesis screens with P elements.
Genetically engineered P elements. The suppressor-P element composite transposon. The P-enhancer trap elements. Cloning of genomic DNA flanking a P element: plasmid rescue and inverse PCR techniques. Protein trap insertion screening.
Gain of function genetics. Targeted ectopic expression by tissue-specific and heat-shock promoters.
Conditional transgene expression approaches using the Gal4/UAS system of S. cerevisiae. Use of the conditional GAL80 allele to regulate GAL4-driven transgene expression. The Gal4/UAS enhancer trap system. The inducible GeneSwitch GAL4 System. The GAL4/UAS based modifier screen.
EP transposable elements and misexpression screens.
Mitotic recombination. The FRT/FLP system of S. cerevisiae to generate genetic mosaics. The FLP-Out cassette technique. Chimeric proteins that are used as markers of recombinant cells.
Mosaic analysis with a repressible cell marker. Mosaic genetic screens.
Reverse genetic approaches to generate loss of function phenotypes. RNA interference-mediated gene silencing. Targeted disruption of gene function by RNA interference. Targeted mutagenesis.

Readings/Bibliography

Reviews and original papers.

Teaching methods

Lectures.

Assessment methods

Oral interview.

Teaching tools

PowerPoint presentations.

Office hours

See the website of Valeria Cavaliere