84893 - GENETICA E GENOMICA FUNZIONALI

Learning outcomes

The student will get deep knowledge about approaches used in forward genetic analysis aimed at functional genetic dissection of complex biological processes. Student will be able to analyze and discuss experimental data from genetic analysis using animal model system and will be able to propose strategies aimed at functional dissection of biological processes of interest using animal model systems.

Course contents

Module 1 - FUNCTIONAL GENETICS

Mutation as main tool of genetic functional analysis aimed at the genetic dissection of a complex biological process.

Description of the classes of loss of function (hypomorphic, amorphic or null) and gain of function (hypermorphic, neomorphic, antimorphic or dominant-negative) alleles and genetic strategies for their characterization.

Genome wide genetic screen as strategy for functional dissection of biological processes.

Main genetic model systems (unicellular: Saccharomyce cerevisiae; invertebrates:Caenorhabditis elegans, Drosophila melanogaster and vertebrates: zebrafish, mouse) and their peculiar methods and use to perform genetic screens alchilant or insertional (retroviral vectors, transposons) mutagenesis.

-Screen to identify alleles that modify (suppress or enhance) a phenotype due to a primary mutation (S. cerevisiae; C. elegans, D. melanogaster, mouse)

-Screen to identify zygotic and maternal lethal mutations (C. elegans, D. melanogaster, zebrafish, mouse)

-Large scale screen (S. cerevisiae; C. elegans)

-Synthetic lethality screen (S. cerevisiae; C. elegans)

-Non-allelic non-complementation screen (C. elegans)

 

Module 2 - FUNCTIONAL GENOMICS

Introduction to microarray based techniques. History, evolution and available technologies. Application of the microarray based technologies to analyse non-coding RNAs and SNPs.

Genome sequencing techniques: from the Genome Project to the Next Generation Sequencing. Basics of the different available NGS platforms.

Comparing NGS versus microarray: applications, potentiality and limits of the different technologies.

Bioinformatics laboratory. Use of free softwares to analyse data of gene expression and gene annotation analysis. Use of the softwares to analyse miRNA and target prediction.

 

 

Readings/Bibliography

PowerPoint presentations in pdf format, reviews and original scientific articles in electronic format.

Reference book:
Philip Meneely
Analisi genetica avanzata.
ed. MacGraw-Hill
anno: 2012
ISBN/EAN: 9788838665196

Teaching methods

Module 1 - FUNCTIONAL GENETICS

The teaching consists of 6CFU: 5CFU of lectures and 1 CFU, corresponding to 15 hours, of practical work in a laboratory. The lectures will be supported by projection of PowerPoint presentations that will be provided to students in pdf format. During the laboratory the students will apply protocols for immunofluorescent detection of molecular markers suitable for the analysis of the phenotypic effect of mutant alleles of a gene important for the development of the nervous system.

 

Module 2 - FUNCTIONAL GENOMICS

The teaching module consists of 2CFU: 1CFU of lectures and 1CFU, corresponding to 12 hours, of practical work in a bioinformatic laboratory. The lectures will be supported by projection of PowerPoint presentations that will be provided to students in pdf format. During the laboratory the students will apply freely-downloaded tools for the design of primers to be used the sequencing genes according to Sanger’s method; practise access to portals for the analysis of data from next-generation sequencing and familirize with tools for target prediction of miRNAs.

Assessment methods

The final exams at the end of the two modules aim at assess the achievement of the learning objectives:

 

Module 1 - FUNCTIONAL GENETICS

- To know the effects on gene function of the main classes of mutant alleles, loss and gain of function, and genetic techniques to discriminate between the different types of alleles.

- To know the main methods used to perform genome wide experimental mutagenesis using the most used animal model systems (invertebrate and vertebrate)

- Know the main approaches of genetic mutational screen to achieve the complete genetic dissection of the molecular mechanism controlling the biological process of interest, in particular methodologies applied to the main genetic models animals (invertebrate and vertebrate)

The final grade for the module 1 - Functional Genetics is defined by the average of the marks obtained in the written discussion of two open-ended questions on topics related to the main objectives of the course.

Module 2 - FUNCTIONAL GENOMICS

- to know the principles associated with the main techniques associated to massive next-generation sequencing and powerfull gene-hybridization processes.

The final grade for the module 2 - Functional Genomics is defined by the average of the marks obtained in the written discussion of two open-ended questions on topics related to the main objectives of the course.

The final mark of the two modules (8 credits) will be the average of the marks obtained in the two modules.

Teaching tools

PowerPoint presentations

Office hours

See the website of Roberto Bernardoni

See the website of Katia Scotlandi