76166 - Molecular Biology

Course Unit Page

Academic Year 2019/2020

Learning outcomes

Define the mechanisms involved in the replication, repair, transcription and translation of information encoded in nucleic acids. Describe the molecular mechanisms that regulate gene expression, including epigenetics. Describe the molecular mechanisms of cell fate determination and differentiation.

Course contents

- Lesson N. 1 - Introduction to Molecular Biology - DNA structure and function

What is molecular biology? The central Dogma. The discovery of DNA as the heritable material. The discovery of the double helix DNA structure. Nucleotides, covalent and hydrogen bonds, DNA orientation. DNA conformations (B, A, Z).

- Lesson N. 2 - Topological properties of DNA and organization of the prokaryotic and eukaryotic genome

Prokaryotic and eukaryotic DNA. Genome size among organisms. Topological properties DNA (supercoiling, linking number, Whriting e Twisting). Topoisomerases I and II: mechanism of actions. The organization of chromatin in nucleosomes. Nucleosome, core particle and chromatosome. Histone proteins. Histone octamer structure, organization and assembly. Levels of DNA packaging: decondensed chromatin, models of condensed chromatin, post-translational modifications of histones, chromosome scaffolds and DNA loops.

- Lesson N. 3 - Basic rules of DNA replication and molecular techniques based on the analysis of DNA replication

Basic rules of DNA replication. Mechanism of DNA replication. Nucleoside analogue incorporation assay in vitro and in vivo for evaluation of cell proliferation. 14C incorporation for evaluation of DNA replication in tissues with very limited renewal. Polymerase chain reaction (PCR): evolution and applications.

- Lesson N. 4 - Site-specific recombinase technologies for gene knock-out/knock-in and lineage tracing techniques

Recombinase-mediated toolbox (Cre-Lox system for gene expression and ablation, inducible recombinases). Lineage tracing techniques to follow the fate of individual cells and their progeny.

- Lesson N. 5 - Molecular mechanisms of stem cell expansion, renewal and differentiation in tissue development, homeostasis and regeneration

Embryonic stem cells (ES). Stem cell symmetric and asymmetric division: stem cell expansion, self-renewal and differentiation. Adult stem cells. Molecular signals to stem cells. Adult tissues with high and low stem cell renewal. Adult stem cell-based therapies for regenerative medicine.

- Lesson N. 6a - Adult stem cell therapies in cardiac regenerative medicine

Leading cause of death: the quest for cardiac regenerative strategies. Adult stem cell therapies for cardiac regeneration. Lineage tracing analyses of the contribution of progenitor cells to the cardiomyocyte lineage during mammalian embryogenesis and regeneration.

- Lesson N. 6b - Molecular basis of cell reprogramming

Cell reprogramming from somatic cells to induced pluripotent stem cells (IPS). Molecular factors for in vitro cell differentiation of induced pluripotent stem cells (IPS) and embryonic stem cells (ES). Applicative aspects of induced pluripotent stem cells (IPS) and embryonic stem cells (ES) in regenerative medicine. Direct reprogramming of somatic cell to specific cell types (transdifferentiation).

- Lesson N. 7a - Cutting-edge technologies for the analysis of DNA replication in stem and mature cells

DNA polymerase proofreading activity. DNA mismatch repair. The immortal strand hypothesis of DNA replication in stem cells. Multi-isotope imaging mass spectrometry (MIMS) for evaluation of DNA replication in stem cells. Combinatorial approaches with MIMS and lineage tracing genetic labelling for studying the source of slowly proliferating tissues.

- Lesson N. 7b - Advanced molecular tools for time-controlled gene expression

Bacterial Tetracycline resistance mechanism. Tet Repressor protein(TetR). Tet-OFF and Tet-ON system for inducible gene expression.

- Lesson N. 8 - Molecular mechanisms of mature cell dedifferentiation

Cell dedifferentiation features and markers. Cell dedifferentiation in tissue regeneration. Growth factor signalling promoting cell dedifferentiation.

All lessons will be held at Aula Magna Patologia Generale, via San Giacomo 14, Bologna

 

Readings/Bibliography

The course is based on the presentation of the material that will be supplied to the students via the "Insegnamenti OnLine - IOL" platform [https://iol.unibo.it/].

Teaching methods

  • Frontal lessons through computer-assisted presentations and critical discussion of scientific articles.
  • Tests at the end of the lessons will be used to support learning.
  • Students will also be progressively led to identify problems of interest and to use telematic tools, such as PubMed Medline, for the deepening of curiosities arising during the lessons.

Attendance to learning activities is mandatory; the minimum attendance requirement to be admitted to the final exam is 60% of lessons. For Integrated Courses (IC), the 60% attendance requirement refers to the total amount of I.C. lessons. Students who fail to meet the minimum attendance requirement will not be admitted to the final exam of the course, and will have to attend relevant classes again during the next academic year.

Professors may authorise excused absences upon receipt of proper justifying documentation, in case of illness or serious reasons. Excused absences do not count against a student’s attendance record to determine their minimum attendance requirement.

Assessment methods

Computer-assisted oral exam: Each student will undergo an individual oral exam. The student will be requested to bring a short presentation (5 slides, 5 minutes presentation) on a topic of choice, among those discussed during the course. The presentation should be prepared by the student in advanced and provided on a USB drive. The student will then be requested to address further issues that may arise from her/his presentation, as well as other topics that have been discussed during the course lessons.

 

Teaching tools

  • Powerpoint presentations
  • Scientific videos
  • Scientific articles and reviews

All the material will be available to the Students via the "Insegnamenti OnLine - IOL" platform [https://iol.unibo.it/].

Office hours

See the website of Gabriele Matteo D'Uva