CELLULAR, MOLECULAR AND THERAPEUTIC TECHNOLOGIES - BIOLOGY, GENOMICS, HUMAN AND MEDICAL GENETICS

Module TECNOLOGIE CELLULARI, MOLECOLARI - BIOLOGIA E GENOMICA

1) Knowledge and understanding

At the end of the course, the student will have to demonstrate knowledge of the theoretical and experimental bases of the main cellular and molecular technologies. The student will have to demonstrate to know in general terms the main technologies for the study of genomics and epigenomics and to apply the latter for the integrated study of the cell and living organisms, with particular enphasis to the Molecular Biomedicine.

 

2) Practical application of acquired knowledge

At the end of the course, the student must be able to identify the essential elements of a biological experiment and hypothesize the best technologies for carrying it out.

The course is organized in 35 hours of lectures with the help of slides and videos and 12 hours of laboratory exercises.

If the course is taught in mixed or distance mode, the necessary changes may be introduced with respect to what was previously stated, in order to comply with the program provided and reported in the syllabus.

Knowledge of the structure and function of cells, genome and cellular epigenome

Attendance of the course is required for at least 50% of the lessons of the module to be admitted to the final exam of the course.

CELL BIOLOGY TECHNIQUES:

- Preparation and life of a cell culture

- cultures of primary and immortalized cell lines

- 2D and 3D cell cultures

- iPSCs technology

- techniques for the transfection of mammalian cells by physical and chemical methods

- general principles of microscopy

 

MOLECULAR TECHNIQUES: · NUCLEIC ACID ANALYSIS AND SEPARATION TECHNIQUES 

- Extraction and purification of nucleic acids from cell lines, biological fluids and biopsies      

- Spectrophotometric techniques for DNA and RNA analysis

 

CLONING VECTORS

- definition and role of the main components of prokaryotic plasmids and eukaryotes

 

TECNICHE DI EDITING GENOMICO

- Zinc‐finger nucleases (ZFNs) (1° generation)

- Transcription activator‐like effector nucleases (TALENs) (2° generation)

- RNA‐guided nucleases that use the clustered regularly interspaced short palindromic repeat (CRISPR) and a conserved CRISPR‐associated protein (Cas) (3° generation)

 

QUANTIFICATION OF GENE EXPRESSION BY PCR AND REAL-TIME PCR

- Low- and high-throughput methods and data analysis

 

QUANTIFICATION OF GENE EXPRESSION BY ddPCR

 

METHODS FOR THE STUDY OF DNA METHYLATION

- Non-specific methods

- Specific methods

- DNA modifications with bisulfite

 

METHODS FOR THE STUDY OF HISTONE CHANGES

- Chip

 

TECHNIQUES FOR THE STUDY OF NON-CODING RNAs

- microRNAs, circRNAs and lncRNAs

- Identification of ncRNAs through in vitro and in silico methods

- ncRNA extraction and analysis

 

INTERACTIONS BETWEEN BIOLOGICAL MACROMOLECULES

- Physical and functional interactions between ncRNAs

- RNA-Protein and protein-protein Interactions

 

BIOINFORMATICS ANALYSIS FOR THE STUDY OF GENOMES AND TRANSCRIPTOMES

 

LABORATORY EXERCISES

 

1.Francesco Amaldi, Piero Benedetti, Graziano Pesole, Paolo Plevani. Techniques and methods for molecular biology. Zanichelli (Italy) 

2.Massimo Romani. Epigenetics. Zanichelli (Italy) 

3.Teaching materials provided by the instructor

 

NOTE: It is necessary to use the most recent edition of the reference texts.

The exam consists of an interview in which a powerpoint will be presented on a topic of scientific literature agreed with the teacher and in which 2 questions will be asked that focus on at least 2 different topics of the program. The exam allows to verify: i) the level of knowledge on advanced cellular and biomolecular technologies; ii) the ability to apply this knowledge to the resolution of specific problems related to the study of Biology and Genomics, with particular reference to Molecular Biomedicine (problem solving and autonomy of judgment); iii) clarity of exposition; iv) the property of scientific language.

The following parameters will be taken into account for the attribution of the final grade:

Grade 29-30 cum laude: the student has an in-depth knowledge of the topics, is able to promptly and correctly integrate and critically analyze the situations presented, independently solving problems even of high complexity; has excellent communication skills and masters scientific language.

Grade 26-28: the student has a good knowledge of the topics, is able to integrate and analyze the situations presented in a critical and linear way, is able to solve complex problems quite autonomously and presents the topics clearly using an appropriate scientific language;

Grade 22-25: the student has a fair knowledge of the topics, even if limited to the main topics; is able to integrate and analyze the situations presented in a critical but not always linear way and exposes the arguments quite clearly with a fair property of language;

Grade 18-21: the student has the minimum knowledge of the topics, has a modest ability to integrate and critically analyze the situations presented and exposes the topics in a sufficiently clear way although the language property is poorly developed;

Exam failed: the student does not have the minimum required knowledge of the main contents of the course. The ability to use specific language is very little or non-existent and is not able to independently apply the knowledge acquired.

 

Learning assessments may also be conducted online, if circumstances require.

To ensure equal opportunities and in compliance with applicable laws, interested students may request a personal interview to plan any compensatory and/or extenuating measures, based on their learning objectives and specific needs. Students may also contact the CInAP (Center for Active and Participatory Integration — Services for Disabilities and/or DSA) contact teacher in their Department (https://www.cinap.unict.it/content/referenti).

Describe the main differences and applications of 2D and 3D cell cultures

Defining genome editing and what its applications can be

Eukaryotic cell transfection techniques and their applications

Main techniques used for the study of the epigenome

Main techniques used for the study of interactions between RNA molecules