12946 - Molecular Biology (L-Z)

Learning outcomes

At the end of the Course, the Student will be able to discuss a number of crucial issues in cellular and molecular biology, including: The nuclear traffic of signal molecules and transcription factors; the main activation pathways of transcription factors and their nuclear action mechanisms with particular reference to the processes of gene expression, cell proliferation and differentiation; the role of epigenetics in the modulation of cellular homeostasis; the molecular mechanisms responsible for stem cell pluripotency.

Course contents

- Introduction to the Molecular Biology field.

What is molecular biology? The central Dogma.

- Molecular mechanisms of stem cell renewal and differentiation in tissue development and homeostasis.

Embryonic and adult stem cells, stem cell potency and differentiation. Adult stem cells in tissue with high cell renewal (i.e. bone marrow, intestine, skin...). Adult stem cells in tissue with low cell renewal (i.e skeletal muscle, brain). Cardiac stem and progenitor cells during embryonic development. Molecular mechanisms of stem cell regulation: extracellular matrix, soluble factors (eg WNT, BMPs, ERBBs receptor ligands ...), communications between adjacent cells and mechanical stimuli. The role of transcription factors in cell differentiation.

- DNA replication - Molecular tools for studying of cell proliferation and renewal based on the analysis of DNA replication.

DNA structure and replication. Nucleoside analogue incorporation assay for evaluation of cell proliferation. BrdU incorporation assay in vitro and in vivo for the evaluation of tissue renewal and regeneration. 14C incorporation for evaluation of DNA replication in tissues with very limited renewal, such as the heart and the brain.

- Advanced molecular tools for gene expression control: Cre/Lox system for gene knock-out/knock-in and lineage tracing analysis and Tet-OFF/Tet-ON system for inducible gene expression.

Gene knock-out/knock-in in animal models by non-homologous recombination, homologous recombination and engineered nuclease. Recombinases. Structure and mechanism of action of Cre recombinase. Cre-Lox system for tissue-specific gene knock-out and knock-in. Inducible recombinases. Lineage tracing analysis to follow the fate of individual cells and their progeny: application examples for the evaluation of the contribution of cardiac progenitors to the generation of cardiomyocytes in cardiac tissue development and homeostasis and following damage. Bacterial Tetracycline resistance mechanism: the "Tet Repressor" protein (TetR). Tet-OFF / Tet-ON system for inducible gene expression.

- Stem cell differentiation, cell reprogramming, transdifferentiation and mature cells dedifferentiation: multiple approaches for tissue regeneration.

Adult stem cell-based therapies for regenerative medicine: examples of success and failure. The regenerative potential of embryonic stem cells. 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). Induced pluripotent stem cells (IPS) and embryonic stem cells (ES) in regenerative medicine. Direct reprogramming of somatic cells into specific cell types (transdifferentiation). Cell dedifferentiation in tissue regeneration.

Readings/Bibliography

The course is based on the presentation of the material that will be supplied to the students via Virtual Learning Environment platform.

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.

Assessment methods

The students will be evaluated by written exam on the EOL - Exam OnLine Platftorm.

The written test consists of 20 multiple-choice questions. Students will receive 1.55 points for each correct answer, 0 points for incorrect or missing answers, and an intermediate score for partially correct answers. The time available to students for the written test is 30 minutes. During the test, the use of support materials, such as textbooks, notes, computer supports, is not allowed. The maximum score obtainable by providing all correct answers is equal to 30 cum laude. The test is considered passed with a minimum score of 18/30.

The final mark of the Integrated Course in Biochemistry will be calculated as weighted average of the marks obtained in the modules of Chemistry and General Biochemistry, Metabolic Biochemistry and Molecular Biology, taking into account the training credits (CFU) of each module.

Teaching tools

• Powerpoint presentations
• Scientific videos
• Scientific articles and reviews

All the material will be available to the Students via Virtual Learning Environment platform.

Office hours

See the website of Gabriele Matteo D'Uva