12946 - Molecular Biology

Learning outcomes

The major outcomes will include the acquirement of modern view on the mechanisms underlying cell growth and differentiation, with particular reference to the modulation of gene expression, epigenetics, nuclear dynamics and signaling. Students will be introduced to the molecular dissection of the action of transcription factors, chromatin remodeling and establishment of tissue specific features. The Course will trace the interplay between cell signaling networks and the changes in genome structure and function. Emphasis will be placed on the discussion of several features of stem cell biology, including the concept of pluripotency, paracrine/autocrine/intracrine regulation, and multilineage commitment. Finally, we will dissect the issue of reprogramming of adult somatic non-stem cells to lineages in which these cells would never otherwise appear. These aspects will form the underpinning for Students' guidance through the newly developing fields of Regenerative Medicine and Precision Medicine.

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 - DNA replication

Basic rules of DNA replication. Control of the initiation of DNA replication. The replicative fork. Proteins involved in DNA replication: helicase, primase, DNA polymerase, sliding camp, single strand binding proteins, topoisomerase, nucleases, DNA ligase, telomerase. Mechanism of DNA replication in leading and lagging strands. Types of DNA polymerases in prokaryotes and eukaryotes. Chemical Mechanism of Chain Elongation.

- Lesson N. 4 - Gene expression

Coding and non-coding DNA. Gene transcription. Coding and template strands. Gene expression regulation: promoters, enhancers, silencers e insulators. RNA polymerase. Initiation, elongation, termination of gene transcription. Maturation of eukaryotic mRNA and alternative splicing. Regulatory levels of gene expression.

- Lesson N. 5 - Advanced molecular tools for gene expression regulation

Recombinases. Cre recombinase structure and mechanism of action. Cre-Lox system for tissue-specific gene ablation and expression. Inducible recombinases. Lineage tracing techniques to follow the fate of individual cells and their progeny. Bacterial Tetracycline resistance mechanism. Tet Repressor protein(TetR). Tet-OFF/Tet-ON system for inducible gene expression.

- Lesson N. 6a - Stem cell renewal and differentiation in tissue development and regeneration

Stem cell potency: totipotent, pluripotent, multipotent stem cells. Stem cell symmetric and asymmetric division: expansion, self-renewal and differentiation. Embryonic stem cells (ES). Adult stem cells in tissue with high cell renewal (i.e. bone marrow, intestine, skin, skeletal muscle, brain). Adult stem cells in tissue with low cell renewal (i.e skeletal muscle, brain). Cardiac stem and progenitor cells during embryonic development. Molecular signals to stem cells.

- Lesson N. 6b - Stem cell based regenerative medicine strategies

Adult stem cell-based therapies for regenerative medicine. The clinical need for cardiac regenerative strategies. Adult stem cell therapies for cardiac regeneration. Lineage tracing analyses of the contribution of cardiac progenitor cells to the cardiomyocyte lineage during mammalian embryogenesis and following cardiac damage. Cell reprogramming from somatic cells to induced pluripotent stem cells (IPS). Molecular factors for in vitro cell differentiation of embryonic stem cells (ES) and induced pluripotent stem cells (IPS). 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. 7 - Molecular mechanisms of cell dedifferentiation 

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

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.

Assessment methods

Computer-assisted oral exam: Each student will undergo an individual oral exam. The student will be requested to bring a short presentation on a topic of choice (5 slides, 5 minutes presentation), 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