Dipartimento di Scienze della Vita e dell'Ambiente - Guida degli insegnamenti (Syllabus)
Knowledge of Biological Chemistry, Molecular Biology and Genetics is required.
The Advanced Molecular Biology module is organized in lectures (5 CFU, 40 hours) and laboratories (1 CFU, 8 hours). Handouts of the lectures, protocols for the laboratories and booking forms for the laboratories are loaded on the Moodle platform of DiSVA website.
Students will acquire knowledge concerning molecular mechanisms involved in the regulation of gene expression at the different levels with a special interest in all post-transcriptional events as well as the most important and most widely experimental approaches used for gene expression analysis.
Ability to apply the knowledge:
Students will be able to apply the acquired knowledge to understand and critically evaluate data in the scientific literature and to perform same of the basic technique for gene expression analysis.
The practical experience during the laboratories together with the discussions on the experimental results will improve student’s autonomy and ability to evaluate critically scientific data. Furthermore, the organization of the laboratories in small groups will facilitate the coordination and communication abilities. The preparation of a Power-point presentation for the final exam will advance the student’s ability to present and discuss scientific literature.
Content (lectures 5 CFU, 40 hours):
Methods for gene expression analysis: northern blotting, RT-PCR, RNase protection. Real-time PCR. DNA microarrays. RNA-Seq. Next-generation sequencing techniques. Reporter genes. Western blotting, proteomic analysis, mass spectrometry, SILAC.
Analysis of DNA and RNA-protein interaction: DNaseI footprinting, chemical probing, cross-linking, ChIP-Seq. Analysis of protein-protein interactions: two-hybrid and three-hybrid systems, GST-pull down. Analysis of translation: cell-free systems, toe-printing, polysomal profiling, Ribo-Seq.
Site-directed mutagenesis. Knock.out and knock-down. CRISPS-Cas system.
The different levels of regulation of gene expression.
Epigenetic modifications: DNA methylation, histone methylation and chromatin remodelling.
Regulation at the post-transcriptional level: RNA binding proteins and RNA binding motifs, mRNA maturation, polyA tail addition, splicing and alternative splicing, mRNA transport, the “post-transcriptional operon” hypothesis, translation, mRNA decay, nonsense-mediated decay, miRNA and siRNA.
Regulation at the post-translational level: protein stability and processing.
Laboratories (1 CFU, 8 hours):
Extraction of RNA from cultured cells; purification of RNA by affinity chromatography and determination of its concentration by spectrophotometric analysis; retro-transcription followed by PCR with primers designed to amplify alternative splicing products; separation of the different amplicons by agarose gel electrophoresis; data analysis.
Methods for assessing learning outcomes:
Evaluation for Advanced Molecular Biology module is performed trough an oral exam that consists of a Power point presentation based on a research article dealing with one of the topics of the program and two questions. In addition, students should present a written report of the laboratory experience.
Criteria for assessing learning outcomes:
Questions aim at verifying the level of knowledge acquired by the students. Power point presentation will show the ability of the student to present and critically discuss the scientific literature and to evaluate experimental data. The laboratory report will show the capacity of the student to understand and perform experiments.
Criteria for measuring learning outcomes:
The level of knowledge acquired by the students is measured with a mark between 0 and 30. In order to pass the exam the final mark must be between 18 and 30. The highest possible mark is 30/30 cum laude.
Criteria for conferring final mark:
Being an integrated course, the final grade is attributed upon evaluation of the overall results obtained in the single modules. Concerning the Advanced Molecular Biology module, the Power-point presentation is evaluated with a mark included between 0 and 24, the answer to the two questions and the laboratory report are evaluated overall with a mark between 0 and 6. 30/30 cum laude is attributed to students particularly able to discuss critically and with great competence about the different topics of both modules.
Biologia molecolare del gene. J.D. Watson, T.A. Baker, S.P. Bell, A. Gann, M. Levine, R. Losick. Casa Editrice Zanichelli. VII edizione. 2015.
Biologia Molecolare. F. Amaldi, P. Benedetti, G. Pesole, P. Plevani. Casa Editrice Ambrosiana. II edizione. 2014.
In addition, review articles from specialized Molecular Biology journals on specific topics are provided during the course.