Learning objectives
Acquiring knowledge and understanding.
The objective is for the students to knowledge over the recent techniques of genome engineering. In particular, regarding the CRISPR-cas9 system and its derivatives. Acquisition of the knowledge for the potential applications of these techniques in the biomedical and biotechnology field. Development of critical sense regarding legal and ethical issues related to the employment of these techniques in particular for human reproduction purposes.
Applying knowledge and understanding.
Through guided analysis of key experiments for the molecular understanding of the development of the science in the field of the genomic engineering, the students will strengthen their competence in addressing the experimental study and application of the techniques of genomic engineering.
Prerequisites
Good knowledge of the structure of nucleic acids, the structure of the genome and the basic mechanisms of duplications, transcription, repair and recombination of DNA.
Course unit content
Genetic manipulation of animals (mammals, Xenopus and zebra fish).
Genetic manipulation of plants and plastids.
Inducible promoters.
Site-specific recombination and conditional mutagenesis.
RNA interference and functional knock down.
Homologous (HDR) and illegitimate (NHEJ) recombination.
The Genome. Short introduction on genome structure, in particular regarding the structure and function of the human genome. The Genomes of lower organisms will be briefly discussed as well.
Recent methods of engineering of genomic sequences. Comparative techniques of genome editing. Zinc Finger Nucleases (ZFNs), TALENs proteins and RGENs ((RNA-guided engineered nucleases). CRISPR Technology. Evolution of the CRISPR technology. Recent advancements for the use of CRISPR technology.
Genomic engineering of human cells. Putative application of these methodologies. Use of the CRISPR technique for studying gene function: disruption of the gene function, insertion, mutation, gene correction and chromosomal rearrangement.
Employment of the “dead-cas9” dCas as versatile and programmable platform to control gene expression and epigenetic regulation. Advancement and development of dCas tools for synthetic biology. Utilizzo del sistema dCas per analisi della trascrizione, effetti epigenetici, microscopia, screening e immunoprecipitazione cromatinica. Employment of dCas-KRAB systems to silence unique and multiple genomic sequences. Advantages and new barriers and prospective of the dCas system as new technological platform.
Potential applications of large-scale genome engineering of different organisms.
Genome editing in regenerative medicine.
Genomic engineering of human reproduction. Ethical and legal aspects. ¬
Full programme
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Bibliography
Reference textbooks No reference textbook. All the material used by the teacher to prepare the lessons will be provided to the students.
Teaching methods
The course consists of lectures on key topics in the program, and in depth focus on topics of particular relevance and interest, with the use of original scientific articles and the assistance of specialist researchers
Assessment methods and criteria
The assessment of the expected learning outcomes is based on a oral exam
with both teachers. Both the knowledge at the molecular level related to the genomic engineering systems
illustrated during the course and the ability to apply this knowledge to the resolution of
experimental problems will be evaluated.
Other information
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