76166 - Molecular Biology

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

- Introduction to the field of Molecular Biology - DNA structure and function

What is molecular biology? The central Dogma of Molecular Biology. 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.

- 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.

- Site-specific recombinase technologies 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: molecular mechanisms and 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.

- Advanced molecular tools for tissue-specific and time-controlled gene expression (Tet-OFF/Tet-ON system)

Basic rules of gene expression. Tet-OFF/Tet-ON system for inducible gene expression.

- Mature cell dedifferentiation: molecular mechanisms and potential therapeutic applications

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

- 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).

- Technologies to control gene expression in humans

Modified RNA.

Readings/Bibliography

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

Teaching methods

• Frontal lessons through computer-assisted presentation 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 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 (I.C.) will be determined as weighted average mark (weights are the relative course credits).

Below the applied formula:

Final score I.C. = [(Cellular Biology and Genetics * 7) + (Cellular Biology and Genetics Laboratory * 1) + (Molecular Biology * 2)] / 10

For example:

Cellular Biology and Genetics (7 credits): 28/30

Cellular Biology and Genetics Laboratory (1 credits): 29/30

Molecular Biology (2 credits): 30/30

Weighted average mark: [(28*7)+(29*1)+(30*2)] / 10 = 28.85

Final mark: 29/30

For more info: https://www.unibo.it/en/teaching/course-timetable-and-exams/about-exams

Teaching tools

All the slides of the lessons are available to the Students via Virtual Learning Environment portal.

Office hours

See the website of Gabriele Matteo D'Uva