Learning objectives
Aims of the course are to know:
- the human genome’s structural and functional features and the pathways involved in its maintenance and expression
- basic knowledge in human genetics, with a focus also on mitochondrial genetics: the hereditary transmission patterns and human haplotypes, and the consequences for human health of mutations in the genome
- research advances in DNA editing, the relationship between the patient's genetic component and the response to drugs and development of potential therapeutic approaches
- the main applications of DNA analysis in the forensic field
- the main experimental models used in biomedical research
- the genetic tools available for functional analysis of genes in the different model organisms
The course aims to:
- to understand how the integration of the different experimental models in vivo and in vitro gives specific answers to the different biological problems
- to acquire the ability to develop and apply experimental strategies for the resolution of genetics and cell biotechnology problems.
Course unit content
The course is divided into two sections that delve into the molecular genetics of humans and model organisms, focusing on the mitochondrial genome and the nuclear genome.
SECTION 1 (in english)
Part 1: THE MITOCHONDRIAL GENOME
- An introduction to mitochondria
- The mitochondrial DNA
- Mitochondrial DNA replication and repair
- Mitochondrial DNA expression
Part 2: MITOCHONDRIAL GENETICS IN HEALTH AND DISEASE
- Mitochondrial genome maternal inheritance, germline segregation and heteroplasmy
- Mitochondrial genome haplogroups and human evolution
- Primary mitochondrial disorders
- Mitochondrial DNA mutations in cancer and aging.
Part 3: EXPERIMENTAL APPROACHES AND THERAPEUTIC DEVELOPMENT
- Model system for the study of mitochondria
- The yeast model system
- Mitochondrial genome editing
- Development of potential therapeutic approaches for mitochondrial dysfunction
SECTION 2 (in italian)
Part 1: HUMAN MOLECULAR GENETICS
- The human genome, the human karyotype
- Genomic and chromosomal mutations and diseases
- Other sources of variability: genetic variants.
- Molecular pathology and identification of disease-associated genes
- Hereditary patterns of monogenic diseases
- Genes and cancer: oncogenes and oncosuppressor genes
- Genetic polymorphisms in forensic genetics and pharmacogenetics
Part 2: LABORATORY ACTIVITIES
Genetic variants analysis
Part 3: MOLECULAR GENETICS OF MODEL SYSTEMS IN VIVO AND IN VITRO FOR BIOMEDICAL AND BIOTECHNOLOGICAL RESEARCH:
- The genome editing in model organisms
- Caenorhabditis elegans, Drosophila melanogaster, Danio rerio, Mus musculus
- Drug discovery
- Cell cultures, iPSCs (Induced pluripotent stem cells)
Full programme
SECTION 1 (in english)
Part 1: THE MITOCHONDRIAL GENOME
- An introduction to mitochondria: mitochondria structure, metabolism and function.
- The mitochondrial DNA: mitochondrial genome structure and sequence, mitochondrial DNA packaging into nucleoids.
- Mitochondrial DNA replication and repair: mitochondrial DNA polymerases, models of mitochondrial DNA replication, mitochondrial DNA mutations, repair and quality control.
- Mitochondrial DNA expression: mitochondrial transcription, RNA maturation, and translation; regulation of mitochondrial gene expression.
Part 2: MITOCHONDRIAL GENETICS IN HEALTH AND DISEASE
- Mitochondrial genome maternal inheritance, germline segregation and heteroplasmy: control of mitochondrial DNA copy number, molecular mechanisms controlling mitochondrial DNA inheritance. Homoplasmy vs heteropplasmy.
- Mitochondrial genome haplogroups and human evolution: definition of haplogroups, haplogroups and environmental adaptation, haplogroups and diseases, the mitochondrial Eve, rates of mitochondrial genome mutations, mitochondrial and nuclear DNA functional interactions.
- Primary mitochondrial disorders: definition, clinical and molecular features, inheritance, tissue specificity. Primary vs secondary mitochondrial dysfunction.
- Mitochondrial DNA mutations in cancer and aging: role of mitochondria in tumorigenesis, mitochondrial dysfunction in cancer, highly oxidative tumors, mitohormesis theory of aging.
Part 3: EXPERIMENTAL APPROACHES AND THERAPEUTIC DEVELOPMENT
- Model system for the study of mitochondria: advantages and disadvantages of model systems commonly used in mitochondrial research.
- The yeast model system: yeast metabolism, yeast genetics, selection and analysis of respiratory mutants, mitochondrial DNA manipulation.
- Mitochondrial genome editing: approaches for mammalian mitochondrial DNA editing (Zn-Fingers, TALENs, mitoARCUS, Deaminases).
- Development of potential therapeutic approaches for mitochondrial dysfunction: gene therapy, mitochondrial gene allotopic expression, heteroplasmic shift, mitochondrial transplantation.
SECTION 2 (in italian)
Part 1: HUMAN MOLECULAR GENETICS
- Human genome: structure and sequence of the human genome, fundamental concepts on cells and human chromosomes, human karyotype.
- Genomic and chromosomal mutations and diseases: alterations in chromosome number, structural alterations and their consequences.
- Other sources of variability: genetic variants.
- Molecular pathology: types of mutant alleles, dominance and recessivity from a molecular point of view. Identification of disease-associated genes: techniques for disease identification. Multifactorial trait genetics.
- Hereditary patterns of monogenic diseases: autosomal recessive, autosomal dominant, X-linked recessive, X-linked dominant, Y-linked and mitochondrial inheritance, complications in Mendelian inheritance.
- Genetic variants in forensic genetics and pharmacogenetics. Forensic Genetics: Genetic markers used in forensic science. Determination of sex, origin of the species, individual profile. Paternity, maternity, familiarity test. Interpretation of the results: statistical considerations. Pharmacogenetics: Genes involved in pharmacokinetics and pharmacodynamics. Polymorphisms in genes that influence the availability of drugs. Genetic polymorphisms. Clinical applications, limitations and future perspectives.
- The two main classes of cancer genes: oncogenes and tumor suppressor genes. Function, type of mutation, activation mechanism.
Part 2: LABORATORY ACTIVITIES
Genomic DNA extraction, genotypic distributions and allelic variants analyses through PCR, digestions with restriction enzymes and electrophoresis on agarose gel.
Part 3: MOLECULAR GENETICS OF IN VIVO AND IN VITRO MODEL SYSTEMS FOR BIOMEDICAL AND BIOTECHNOLOGICAL RESEARCH
- Genome editing on model organisms: CRISPRCas9.
- Caenorhabditis elegans: Genetic analysis. Functional analysis by RNAi. Genome-wide screens application for the study of human pathologies.
- Drosophila melanogaster: Genetic techniques. Methods based on transposable elments. Role of Drosophila in drug discovery and as a model in the study of human pathologies.
- Danio rerio, Zebrafish: Life cycle. The production of homozygous haploid embryos. Morpholino tecnique. Knock out of genes by Zn finger nucleases, TALENs and CRISPRs-Cas9. Zebrafish as model organism for the study of human pathologies.
- Mus musculus, mouse: Transgenesis. Knock-out and knock in of genes.
- Drug discovery in model organisms.
- Cell cultures: primary, stabilized and immortalized cell lines, iPSCs (Induced pluripotent stem cells).
Bibliography
- Philip Meneely, Analisi Genetica Avanzata
ED. McGraw-Hill
- T. Strachan, A.P.Read, Genetica molecolare umana, Zanichelli
- Mitochondria I. E. Scheffler. Wiley
- The Human Mitochondrial Genome, G. Gasparre, AM. Porcelli, Elsevier
- Ricci, Previderè, Fattorini, Corradi
-La prova del DNA per la ricerca della verità, Giuffrè Ed
- Original papers suggested by the professor
Teaching methods
Classes will combine traditional frontal lectures, in which topics will be presented by the instructor, with data analysis and discussion of recent relevant scientific literature, where the students will be asked to actively participate. Students will be required to read research articles, which will be discussed during class. All activities in section 1 will be conducted in English.
The second section of the course also includes laboratory activities.
The teaching materials used in the lessons and any other useful resources will be provided to the students through the Elly platform. To access these online resources, enrollment in the course is required.
Assessment methods and criteria
The acquired knowledge will be assessed with a single written test at the end of the two sections, which will include: a multiple-choice test on the topics covered in section 1 and a written test in which the student will have to answer two open-ended questions related to the topics covered in section 2. The test will last 2.5 hours.
In this test the knowledge and understanding of the different topics covered during the lessons and the ability to communicate clearly and with language properties, ideas and concepts will be assessed.
