The last 10 years have seen a revolution in the possibility to study the regulation of DNA metabolism at the single cell and single molecule level in live cells. Sophisticated technologies allow us to quantify movements, interactions and dynamics of the actors impacting on genome biology. The course of Advanced Molecular Biology aims at providing a deep overview of the mechanisms that regulate cellular processes in complex organisms in health and disease. In particular, mechanisms controlling cell division, proliferation and quiescence are integrated with processes required to maintain the integrity of the genome; on the other hand, cell plasticity and adaptation are required for cellular homeostasis and response to environment. The input comes from the genome; genome structure and function maintain cell identity and regulate transcription through a plethora of different and integrated epigenetic mechanisms. Genome integrity, transcription, tridimensional organization and compartmentalization in the nuclear space are quantitative and dynamic parameters that, occurring at single cell level, govern tissue organization and specialization. These topics will be discussed in model organism and in mammalian cells, with a particular focus on human pathologies, especially cancer, linked to alterations in the molecular mechanisms of these processes. Students will also become familiar with advanced approaches in molecular biology and cutting edge biophysical methodologies to obtain quantitative data on proteins-proteins, protein-DNA interactions, reversible post-translational modifications, nucleic acid processing.
Risultati apprendimento attesi
After this course, the student will be: -familiar with the mechanisms underlying cell proliferation, differentiation and its regulatory circuits -experts in key transcriptional networks that control cell growth, proliferation, DNA repair and their alteration in cancer -familiar with the molecular mechanisms responsible for preserving genome integrity and their role in counteracting tumorigenesis -familiar with the epigenetic layers that define a specific epigenome and their possible alteration in diseases -experts in the methodologies and cutting-edge technologies to study single molecules and their interactions, and to interpret protein networks involved in cancer -able to critically evaluate the advantages and disadvantages of model systems to study molecular processes - design the best experimental strategies to answer a specific scientific question; - critically evaluate hypotheses and working models
Periodo: Secondo semestre
(se vi sono più edizioni controllare anche il periodo dell'edizione, che può essere diverso)
- The genome organization in eukaryotic cells - Non-coding RNAs - Epigenetic regulation of gene expression (DNA methylation, histone variants and modifications, chromatin accessibility and ncRNAs association, etc), technologies and alterations in human diseases - Tridimensional organization of the genome (polymer folding principles, TADs and long-range interactions), technologies and alterations in human diseases - Genome regulation through compartmentalization and phase separation (nucleolus, transcription factories and repression bodies, LADs, nuclear pore, etc), technologies and alterations in human diseases - Models for studying genome organization (CRISPRs/cas9 cell systems, organoids, mouse models etc) - Control of cell proliferation and growth - The cell cycle control systems (S phase, mitosis, completion of mitosis and cell division) - DNA repair and DNA damage tolerance mechanisms - Single cell analysis of transcription and genome structure - Single molecule tracking to study transcription factors consensus site recognition and other proteins involved in DNA metabolism - Super-resolution imaging approaches to study the localization of transcription factors, repair protein and other DNA metabolic factors and accessible regions of the genome. - Molecular genetics approaches to study protein/DNA, protein/protein and protein/RNA interactions at the single molecule resolution. - In vivo imaging to study key DNA metabolic factors mediating cancer cell proliferation and metastasis - Pathologies related to increased genome instability - Cell response to environmental cues (infection and immune response, heat shock genes response, vernalization, mechanical stress, tumor microenvironment, etc).
The course will require basic knowledge of cell biology and biochemistry and a good knowledge of molecular biology and genetics. Students must be aware of the principles of the DNA metabolisms, notably transcription, replication, repair and recombination. They must have knowledege of concepts related to cell growth, cell cycle progression, cell differentiation, and basic knowledged of formation of organs and tissues. They must be aware of concepts related to multi-subunit protein complexes, and on the principles of enzymatic activities.
Traditional lectures supported by slides integrated by on-site interactive discussion sessions, in which student's involvement will be promoted through assignment of scientific papers in advance, with the purpose of stimulating group discussions and to develop students' critical and communication skills. All the teaching materials will be available through the Ariel platform. Regular attendance is strongly suggested.
Materiale di riferimento
The topics covering most of the course are very novel and as a consequence there is no comprehensive text. Web sites and review papers will be indicated to the students. Lectures will be recorded made available through the Teams and Ariel platforms, together all the materials instrumental to allow the students to participate to the on-site discussion lessons, such as original articles and reviews.
Modalità di verifica dell’apprendimento e criteri di valutazione
Learning assessment will be through an oral exam at the end of the course. Examples of exams will be presented during the course.