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

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

- DNA Structure, function and regulation: from discovery to modern concepts

The discovery of DNA as the heritable material. The discovery of the double helix DNA structure. Overview of DNA manipulation, the birth of biotechnology, gene regulation, genomics, precision and systems 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.

- DNA- and RNA-based tools to control gene expression.

DNA plasmids. Viral vectors (retroviruses, lentiviruses, adenoviruses, adeno-associated viruses – AAVs). RNA-based approaches (RNAi, shRNA, miRNA and antagomir and siRNA, modified mRNA).

- 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 MORPHOFUNCTIONAL SCIENCES 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, 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.

Students with learning disorders and\or temporary or permanent disabilities: please, contact the office responsible (https://site.unibo.it/studenti-con-disabilita-e-dsa/en/for-students) as soon as possible so that they can propose acceptable adjustments. The request for adaptation must be submitted in advance (15 days before the exam date) to the lecturer, who will assess the appropriateness of the adjustments, taking into account the teaching objectives.

Teaching tools

All educational materials presented will be available to students through Virtual Learning Environment platform [https://virtuale.unibo.it/] .

Office hours

See the website of Gabriele Matteo D'Uva