AIMS AND LEARNING OUTCOMES

The course has the aim to build in the student of the medical school a detailed and critical knowledge of the molecular biology of the nuclear processes in the higher Metazoa. This should help in understanding that the malfunctioning of these delicate mechanisms is at the basis of numerous diseases. Furthermore, the student will develop competence in the most used molecular investigation techniques in the biomedical research laboratories. All the acquired concepts have the aim to develop in the student specific abilities in the acknowledgement of how diseases arise from molecular imbalances which bases, in the majority of cases, reside in the alteration of gene expression control, both due to genetic as well as epigenetic causes.

TEACHING METHODS

The course consists of 20 hours of classroom training with theoretical lessons on all topics of the program. Students will also be engaged in discussions of the main topics of the course.

SYLLABUS/CONTENT

Genome comparative description: gene organization and distribution, multi-gene families, repetitive sequences, unequal crossing over, exon shuffling, transposons, pseudogenes.

DNA mismatch repair. Direct reversal. Nucleotide excision repair, base excision repair, double strand break repair and diseases arising from their defect. Molecular mechanisms of homologous recombination and non-homologous end joining in DNA double strand break repair.

Recombinant DNA. Plasmids and restriction endonucleases. DNA Ligase. Prokaryotic transformation. Genomic Libraries and screening assays.

Nucleic acid hybridization and its employ in molecular diagnostics.

DNA polymerase chain reaction (PCR), real time PCR and its employ in diagnostics. cDNA retrotranscription, DNA sequencing.

Transgenic animals production via microinjection and knock out method

Organization ad regulation of gene expression. The eukaryotic transcription complex. Transcription factors, promoters, enhancers, insulators and Locus Control Regions (LCR).

Schemes of histone modifications, maintenance and propagation. DNA methylation. Epigenetics mechanisms in cancer.

Alternative splicing, RNA editing, messenger nuclear exportation and coherence control, RNA interference, microRNA and regulation of mRNA stability.

The birth of induced pluripotent stem cells: methods of production and use in biomedicine and cellular therapies.

Gene therapy: choice of candidate diseases, in vivo and ex vivo transfection techniques, viral and non-viral vectors, DNA vaccines. Most used clinical protocols all over the world.

RECOMMENDED READING/BIBLIOGRAPHY

Alberts B, Johnson A, Lewis J et al. Biologia Molecolare della cellula, VI Ed, Zanichelli