ADVANCED MOLECULAR BIOLOGY

This is an advanced course of molecular biology that aims to broaden the knowledge in the field of pharmaceutical biotechnology. In particular the molecular mechanisms of several pathologies and the experiments planned to identify specific disease markers will be elucidated. The molecular mechanism of some drugs able “to cure” defective genes (for example by promoting exon skipping) will also be exposed.

-Basic Molecular Biology: a brief review on macromolecules and macromolecule interactions; Gene structure and gene transcription; Regulation of gene expression
-Molecular tools for studying genes and gene activity, mutagenesis techniques, genome editing by CRISPR-Cas9 system, zinc finger nuclease,TAL effector nuclease
-Non coding RNAs, microRNA: expression, processing, mechanism of action of miRNA; miRNA and interfering RNA (RNAi); RNAi as tool to study gene function; RNAi and therapeutic perspectives
-Molecular basis of cancer (brief summary); oncogenes and oncosuppressors; cancer and epigenetic; miRNAs and cancer; molecular therapy of cancer, immunological therapy of cancer;
-Molecular basis of Duchenne muscular dystrophy and cystic fibrosis (brief summary): frame shift and non sense mutations;
Disease therapies acting on transcript maturation:exon skipping, ODN therapy for correct reading frame rescue; efficiency of ODN delivery; disease therapies acting on translation: stop codon read through.

Finally, a practical course is planned in which GFP expression will be knock down by RNA interference. Students will utilize eukaryotic cell lines, protein and shRNA expression vectors, , analytical methods such as western blotting, cytofluorimetry, immunofluoresce-confocal microscopy