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

- 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.

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

Basic rules of DNA 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.

- Control of gene expression - Advanced molecular tools for tissue-specific gene expression regulation: Tet-OFF/Tet-ON system

Basic rules of gene expression. Technologies to control gene expression in humans. Modified RNA. Tet-OFF/Tet-ON system for inducible gene expression.

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

Embryonic and adult stem cells, stem cell potency and differentiation. Adult stem cells in tissues with high cell renewal (i.e. bone marrow, intestine, skin...). Adult stem cells in tissues 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, communications between adjacent cells and mechanical stimuli. The role of transcription factors in cell differentiation.

- DNA damage, repair and recombination - Endonucleases and gene editing

- Technologies based on site-specific recombinases for tissue-specific gene knock-out/knock-in and "lineage tracing" analysis (Cre/Lox system)

Cre-Lox system for tissue-specific gene knock-out/knock-in. Lineage tracing analysis to follow the fate of specific cell populations and their progeny: basic concepts and application examples for the evaluation of the contribution of specific cell populations during tissue regeneration.

- Differentiation of adult stem cells: therapeutic applications

Adult stem cell-based therapies for regenerative medicine. The controversy of adult stem cell therapies for cardiac regeneration: the emerging consensus based on lineage tracing analyses.

- 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).

- Cell dedifferentiation: molecular mechanisms and potential therapeutic applications

Cell dedifferentiation features and markers. Cell signalling pathways in cell dedifferentiation. Cell dedifferentiation in tissue regeneration.

Readings/Bibliography

Iwasa e Marshall - Biologia Cellulare e Molecolare di Karp

In alternativa è possibile utilizzare qualsiasi libro di testo di biologia molecolare, in italiano o in inglese, ad esempio:

Krebs et al. - Lewin's Essential GENES

Clark et al - Molecular Biology

Lodish et al - Molecular Cell Biology

Watson et al. - Molecular biology of the gene

Krebs et al. - Lewin's GENES XII

Albert et al. - Molecular Biology Of The Cell

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.
  

Attendance to this learning activity 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 authorize 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

The students will be evaluated by written exam.

The written test consists of 31 multiple-choice questions (15 questions for each module plus 1 questions randomly selected from one of the two modules), with only one correct answer per question. Students will receive 1 point for each correct answer, and 0 points for incorrect or missing 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 Applied Biology and Molecular Biology taking into account the training credits (CFU) of each module.

Teaching tools

All educational materials presented will be available to students through Virtual Learning Environment platform.

 

Office hours

See the website of Gabriele Matteo D'Uva

See the website of Carmen Miano