3. CRISPR WARS: The Gene Strikes Back

Luis Povoas (Gene Translation Laboratory)

Some years ago, during the process of constructing a fly model to study human diseases our lab found a very interesting protein that seems to be responsible for many different processes in the cell. This protein is a derivate of a group of proteins responsible to introduce amino acids in tRNA, to enable protein synthesis. We called it SLIMP (seryl-tRNA synthetase-like insect mitochondrial protein). Past studies suggest that this new protein has acquired an essential function in insects, being one of the most interesting results obtained so far, the role of SLIMP in cell cycle progression. To uncover more of SLIMP’s functions, our lab is using a revolutionary technology that changed the way we make science. This technology is the CRISPR editing system. This tool enables us to precisely edit the genome of live cells. With it, we can remove genes (knock-out), introduce genes (knock-in), introduce markers to follow a specific protein (tags) and others. Therefore, with this technology, we expect to uncover SLIMP intracellular location, protein-protein interactions and try to understand the SLIMP knockdown phenotypes in relation to DNA damage and cell cycle checkpoints. During this workshop, you will be introduced to concepts of molecular cloning, cell culture and CRISPR technology. May the science be with you!

One of the main hallmarks of ageing is cellular senescence, the phenomenon by which normal cells stop dividing. Senescent cells accumulate in an organism over time, secreting pro-inflammatory molecules and contributing to age-related diseases. There is an increasing interest in clinical medicine to better identify and target senescent cells, as their elimination may delay and ameliorate some age-associated diseases. 

This course provides hands-on experience using different techniques to induce cellular senescence in normal cells. We will learn state-of-the-art techniques to study the molecular biology of senescent cells, analysing in vitro and in vivo samples. Finally, we will perform classical protocols, such as SAβgal staining, to detect senescent cells. These techniques will help us to understand, identify and target senescent cells for clearance, which is a promising therapeutic approach to extend health span.

4. An omics complex puzzle? Let’s decipher it by using Bioinformatics!

Olfat Khannous (Molecular Modelling and Bioinformatics) & Marina Murillo (Gene Translation Laboratory)

Complex biological diseases such as cancer and Alzheimer are caused by a combination of genetic and environmental factors, and in many cases we have not yet identified or established a relationship between them to shed light to the understanding of the origin and progression of such biological problems.

Multi-omics profiling and integration combines different types of data for a better understanding of these complex diseases. 

One of the branches of the ‘Omics’ sciences is transcriptomics which involves the study of all the RNA molecules (including mRNA, tRNA, rRNA, and other non-coding RNAs) within a cell, otherwise known as the transcriptome. Consequently, by analyzing the transcriptome researchers can determine if a gene is turned on or off in the cells and tissues of an organism.

On the other hand, metagenomics is the study of the genetic material (genomes) from a mixed community of organisms of a sample. It allows us to describe which microbial communities are living within each individual (microbiome) and its relationship with host genetics, diet and environment can reveal links with disease or health.

In our workshop you will be immersed in the computational world (dry lab) understanding its synergy with the experimental lab (wet lab), and you will learn how to obtain transcriptomics and metagenomics data and also how to use bioinformatics tools to analyze them and to add missing pieces to these mentioned complex puzzles.

5. CRISPR/Cas9 to generate a KO

Camilla Bertani (Translational Control of Cell Cycle and Differentiation)

My laboratory is focused on studying a family of proteins called Cytoplasmic Polyadenylation Element Binding Proteins (CPEBs), are sequence-specific mRNA-binding proteins that control translation in development, health and diseases such as cancers. To evaluate the role these proteins, play in cancer cells, a good approach is to induce KO of the protein and evaluate the resulting phenotype.

For this reason, in the laboratory we have designed a possible genomic editing approach, based on the CRIPR / Cas9 technique. Our aim is to induce the deletion of an exon present in the genomic sequence of these proteins, in order to block their functionality.

Therefore, before being able to evaluate the phenotype, it is necessary to evaluate which of the clones we have obtained really presents the deletion of the exon and also, to evaluate if in homozygous or heterozygous form.

For this reason, it is essential to conduct the screening of clones, in particular the analysis of genomic DNA, through the use of a technique called PCR.

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