66619 - Molecular Genetics

Learning outcomes

Students will acquire notions about the molecular genetics of model systems such as M. musculus and H. sapiens and the molecular methods to analyze DNA and RNA and generate mutants through gene inactivation and transgenesis.

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

See the website of Giorgio Milazzo