B9729 - BIOLOGIA MOLECOLARE (LZ)

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

The course is part of the Integrated Course in Biology and Genetics.
At the end of the Integrated Course in Biology and Genetics, the student will have acquired essential theoretical knowledge of the fundamental principles of cell and molecular biology and genetics, as well as their applications in the biomedical field.

SPECIFIC CONTENTS OF THE COURSE:

- Introduction to Molecular Biology

What is Molecular Biology. The central dogma of Molecular Biology.

- Embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs): molecular mechanisms and therapeutic applications

Stem cell potency. Embryonic stem cells. Cell differentiation and the role of transcription factors. Examples of embryonic stem cell differentiation into specific cell types and related therapeutic applications. Cellular reprogramming and induced pluripotent stem cells (iPSCs). Examples of potential therapeutic applications based on iPSCs. Direct reprogramming into specific cell types.

- Molecular techniques for the study of cell proliferation and renewal based on DNA replication analysis

Basic principles of DNA replication. Nucleoside analogue incorporation assays for the evaluation of cell proliferation. In vitro and in vivo BrdU incorporation assays for the study of tissue regeneration. Carbon 14 incorporation for the assessment of DNA replication in tissues with limited cell turnover, such as the brain and heart.

- Conventional gene editing in preclinical models: knock-out and knock-in strategies

Gene knock-out and knock-in in animal models through non-homologous recombination, homologous recombination, and engineered nucleases.

- Conditional gene editing in preclinical models and lineage tracing analysis

Recombinases. Structure and mechanism of action of Cre recombinase. The Cre-Lox system for tissue-specific gene knock-out and knock-in. Inducible recombinases for the temporal control of gene knock-out and knock-in. Lineage tracing analysis to follow the fate of individual cells and their progeny: basic concepts and representative applications for evaluating the contribution of specific cell populations during tissue regeneration. The Tet Repressor (TetR) protein derived from the bacterial tetracycline resistance mechanism. Tet-OFF/Tet-ON systems for inducible gene expression.

- Differentiation of adult stem cells: molecular mechanisms and therapeutic applications

Adult stem cells in tissues with high cell turnover (e.g. sangue, intestine, skin, cornea) and regenerative medicine therapies based on their use, including in combination with gene therapy. Adult stem cells in tissues with low cell turnover (e.g. skeletal muscle). The controversy surrounding adult stem cell therapies for cardiac regeneration: emerging consensus based on lineage tracing analyses. Neural stem cells and their potential modulation in regenerative medicine strategies.

- Dedifferentiation of mature cells: molecular mechanisms and potential therapeutic applications

Features of cellular dedifferentiation. An example of cellular dedifferentiation or stem cell differentiation depending on developmental stage: limb regeneration in salamanders. Cellular dedifferentiation in liver regeneration. Partial cellular dedifferentiation during heart regeneration in non-mammalian vertebrates and neonatal mammals. Potential strategies for cardiac regeneration based on the stimulation of cellular dedifferentiation and cell proliferation.

Readings/Bibliography

All teaching materials presented during the lectures will be made available to students through the Virtual Learning Environment platform [https://virtuale.unibo.it/ ].

For further study, students are free to use any Molecular Biology textbook, in either Italian or English. By way of example, the following volumes are recommended:

Iwasa e Marshall - Biologia Cellulare e Molecolare di Karp

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

Each block of the Integrated Course (I.C.) in Biology and Genetics will receive an evaluation and the final grade will be determined collectively by the teaching staff, taking the weighted average into account.

As for the Molecular Biology module, students will be assessed through a written exam.

The written exam consists of 31 multiple-choice questions, each with only one correct answer. Students will receive +1 point for each correct answer and 0 points for each incorrect or missing answer.

The time available for the written exam is 30 minutes. The use of any supporting materials, such as textbooks, notes, or electronic devices, is not permitted during the exam. The maximum achievable score, obtained by answering all questions correctly, is 30 with honors. The exam is considered passed with a minimum score of 18/30.

Students with learning disabilities or temporary/permanent impairments are encouraged to contact the dedicated office (https://site.unibo.it/studenti-con-disabilita-e-dsa/it/per-studenti ) as early as possible in order to agree on appropriate compensatory measures. The request must be submitted in advance (15 days before the exam date) to the instructor, who will assess the suitability of the measures in light of the learning objectives.

Teaching tools

All the material will be available to the Students via Virtual Learning Environment platform [https://virtuale.unibo.it/ ]

Office hours

See the website of Gabriele Matteo D'Uva