Pharmacy

The Molecular Biology course examines aspects related to structure, organization, replication and repair of the genetic material, as well as the flow of genetic information through transcription and translation. Mechanisms underlying RNA maturation and control of gene expression at different levels (transcriptional, post-transcriptional, translational and post-translational) are illustrated, also analyzing the role of non-coding RNAs. Basic methods for the analysis of nucleic acids and the study of gene expression are discussed. Finally, some translational aspects of pharmacological interest of RNA interference and genome editing are addressed. Detailed program of the course follows. COMPOSITION, STRUCTURE AND PROPERTIES OF NUCLEIC ACIDS. [4 hours] Bases, nucleosides and nucleotides. DNA B, A, Z and alternative structures. Properties of DNA: absorbance, denaturation, renaturation, hyperchromic effect and melting temperature (Tm). RNA: modified bases and main differences between RNA and DNA. Secondary and tertiary structures. Catalytic RNAs. RNA viruses. DNA topology: supercoiling and topoisomerases. Organization of bacterial chromosome, chromatin and eukaryotic chromosomes. Nucleosomes, histones and histone code. Mitochondrial genome. DNA REPLICATION AND REPAIR. [6 hours] DNA duplication: mechanism and origins of replication. DNA replication in prokaryotes and factors involved. DNA polymerases (fidelity and processivity). DNA replication in eukaryotes. Telomeres and telomerase. DNA replication and cell cycle. DNA damage: spontaneous, chemical and physical damages. Repair mechanisms: mismatch repair (MMR), direct and excision repair (BER, NER). Double strand break repair (HR, NHEJ). Cellular responses to DNA damage. TRANSCRIPTION AND TRANSCRIPTIONAL REGULATION. [18 hours] General features and RNA polymerases. Transcription in prokaryotes: initiation (promoters and sigma factors) and termination. The lac operon: lac repressor and CAP protein. The trp operon: attenuation. Transcription and transcriptional regulation in eukaryotes. RNA polymerases and eukaryotic promoters. RNA polymerase II: promoters, basal transcription factors and transcriptional initiation. Elongation and termination. Enhancers and silencers. DNA binding motifs of transcription factors: helix-turn-helix, zinc finger, leucine zipper, helix-loop-helix. Signal transduction and transcriptional activation. DNA methylation and CpG islands. Histone modification and chromatin remodeling. Maturation of rRNA and tRNA in prokaryotes and eukaryotes. Eukaryotic mRNA processing: capping, splicing and polyadenylation. The alternative splicing. Trans splicing. Splicing defects as source of human pathologies and as theraputic target. RNA editing. GENETIC CODE, TRANSLATION AND TRANSLATIONAL REGULATION. [6 hours] The genetic code: deciphering and characteristics. Open reading frame (ORF), reading frame, mutations and their consequences. TRANSLATION: the machinery (mRNA, tRNA, ribosomes and aminoacyl-tRNA synthase). Translational start in prokaryotes and eukaryotes. Translational elongation and termination. TRANSLATIONAL REGULATION: global and transcript-specific control mechanisms (eIF2alpha, ferritin, uORFs and GCN4, cytoplasmic polyadenylation). POST-TRANSCRIPTIONAL CONTROL AND REGULATORY RNA. [8 hours] RNA degradation and regulation of RNA stability (AU-rich elements). Regulatory RNA in prokaryotes: riboswitches and CRISPR. Regulatory RNA in eukaryotes. MicroRNAs: biogenesis, processing and functions. siRNAs and RNA interference (RNAi). Long noncoding RNAs (lncRNAs). Circular RNAs (circRNAs). Applications: gene-specific inhibition by antisense technology and RNA interference. Genome editing, zinc finger and talen nuclease, CRISPR-Cas9. TECHNIQUES. [6 hours] The recombinant DNA technology (restriction enzymes, cloning vectors, genomic and cDNA libraries, expression vectors and recombinant proteins). Basic methodologies for the study of DNA and gene expression: Southern and Northern blot, colony hybridization, microarrays. DNA sequencing: Sanger method and next generation sequencing (NGS). PCR, Real Time PCR and applications. DNA fingerprinting. Techniques for studying DNA-protein interactions: Electrophoretic Mobility Shift Assay (EMSA), DNA footprinting, chromatin immunoprecipitation (ChIP).

For an adequate learning, it is essential that at the beginning of the course the student possesses sufficient knowledge of Biology and General Chemistry as provided by the Pharmaceutical Biology and General and Inorganic Chemistry courses delivered during the first year of the study programme. It is also important and desirable that the student has acquired basic elements of Organic Chemistry and Biochemistry, with particular regard to the main structural and functional properties of proteins.

Any university-level textbook of Molecular Biology covering the syllabus can be considered adequate, and the student can decide on the basis of his preferences which manual seems clearer and more responsive to his needs. Having said so, her below are some recent textbooks where the topics covered in class can be found: 1. Amaldi A. et al. BIOLOGIA MOLECOLARE Terza Edizione Casa Editrice Ambrosiana 2018 2. Watson J.D. et al. BIOLOGIA MOLECOLARE DEL GENE Ottava Edizione Zanichelli 2022 3. Additional/supplementary material made available by the teacher either on the e-learning or Classroom platforms.

Lesson attendance is strongly recommended. Attendance aims to both facilitate the learning process and outline the knowledge level required at the exam. Students unable to attend the lessons due to force majeure can take advantage of the detailed program, of the teaching materials eventually made available and receive assistance from the teacher.

Evaluation takes please through an oral exam typically consisting of 3-4 questions aimed at assessing the knowledge acquired by the student on the various topics covered during the course. To pass the Molecular Biology exam, the student must show to have acquired key competences on structure and functions of nucleic acids, on the mechanisms regulating gene expression, including the role of non-coding RNAs, and to understand the implications, relevance, and possible applications of this knowledge in the pharmacological field. To achieve the highest marks (30/30 cum laude), the student must show excellent command of the various topics covered during the course as well as the ability to connect them in a logical and coherent way.

The Molecular Biology course consists in classroom lectures. The different topics covered during the course can be easily found on the recommended textbooks. Auxiliary teaching material may be made available to students on the e-learning website and/or by Classroom. The teacher remains available for clarification on specific topics upon appointment.