08664 - GENETICA MOLECOLARE

Learning outcomes

At the end of the course, the student possesses in-depth knowledge in the fields of molecular genetics and genomics, related to various model organisms such as yeast, Arabidopsis, Drosophila, C. elegans, mouse, and human. In particular, the student is able to:Analyze and confidently discuss topics in molecular genetics, focusing primarily on the mechanisms of genetic material organization in model organisms, nucleic acid–protein interactions, and the structural and functional complexity of genomes, with particular attention to the regulatory mechanisms of gene expression (transcription and splicing);Understand the strategies, methods, and technologies used to address questions typical of molecular genetics and genomics;Learn to apply the acquired knowledge to the resolution of specific problems (problem solving).

Course contents

Gene interaction: Recessivity, dominance, suppression, enhancement/synthetic lethality.
Complementation test and its exceptions.

Methods for nucleic acid sequencing: DNA/RNA for the study of gene function

Sanger, automated sequencing, capillary electrophoresis sequencing, NGS (Illumina), SMRT (Single Molecule Real Time sequencing), Nanopore, SLAM (Thiol(SH)-linked Alkylation for the Metabolic sequencing of RNA).

Methods to analyse  modifications of DNA and chromatin (Methyl-ChIP, ChIP-seq, ATAC-seq, 3C, 4C, Hi-C) for:

1. defining the methylation state of the genome (methylome),

2. genome-wide mapping of a transcription factor,

3. distribution of histone modifications across the genome and their functional significance,

4. organization of chromatin architecture in the interphase nucleus.

Saturation and targeted mutagenesis methods applied to model organisms such as Mus musculus.

Embryonic stem cells: experiments by Martin Evans.

Knock-out technologies: Capecchi method; CRE/LOXP method; CRISPR/CAS9 methods, base-editing, Prime editing, TALEN.

Knock-down technologies applied to mammalian cells: siRNA, shRNA, CAS13, dCAS9-KRAB-MECP2.

Transgenic technologies: random integration of minigenes, targeted integration, use of inducible systems for transcriptional activation (GAL4-UAS system, Tet on/off, RU486, dCAS9-VPR).

Cancer Genetics. Viral oncogenes, cellular oncogenes, tumor suppressor genes, mutator genes.

Genetic screenings:
Planning of genetic strategies for the identification of new gene functions: genetic screenings using shRNA and sgRNA lentiviral libraries to identify new factors involved in oncogenesis

Readings/Bibliography

Suggested:

5th Edition

Human Molecular Genetics By Tom Strachan

ISBN 9780815345893, 784 Pages, 535 Color Illustrations

Published December 6, 2018 by Garland Science

Teaching methods

Lecture with PowerPoint slides

 Reading of a couple of scientific articles

Assessment methods

40 multiple-choice questions

3 open-ended questions

2 problems taken from the scientific literature

1 experimental design question

Teaching tools

Slides and articles provided by the professor

Office hours

See the website of Giovanni Perini