Researchers of the Department of Biochemistry and Molecular Biology are studying the validity of massive DNA sequencing technologies for the analysis of gene expression in organisms, which allows to know which gene is active at each moment and at which level of intensity. As a model for this study, scientists chose to use Saccharomyces cerevisiae yeast, an organism commonly used in basic research and in biotechnology. The information obtained from this organism can be applied to research on living beings.
The development of the first effective methods to obtain DNA sequencing just over 30 years ago represented a revolution which has since then determined how biology has evolved. Progress in this field has resulted in everything from identifying mutations associated to certain diseases to the complete elucidation of genomes of complex organisms and even human beings. Nevertheless, despite the evolution technology has experienced in the past 30 years, the foundations of biology continue unchanged and this clearly limits the possibility of further progress. This scene has changed drastically in the past few years, with the advent of new technologies in DNA sequencing known by different names, one of which is "massive parallel sequencing".
Although each technique is based on different processes they all hold a common trait which is the capturing of hundreds of thousands of independent sequences at unsuspected speed and without the need for complex sample processing. This has made it possible to explore massively the genetic diversity of microbe communities, to increase largely the number of species in the catalogue of known genomes and to consider sequencing the genomes of individual humans, such as those of the accoladed James Watson.
The potential of these technologies does not stop with the sequencing of genomes; their enormous capacity to generate data can aid in the assessment of genome activities, i.e. the expressions of genes at a specific moment - lack of nutrients, changes in temperature, being exposed to toxic agents - and their level of intensity through the quantification of messenger RNA molecules (mRNA) present in a cell or tissue (sequencing of RNA). This is possible with a simple in vitro transformation of the messenger RNA into DNA - known as cDNA - and obtaining the sequence of millions of molecules of this cDNA. Essentially, the larger the number of sequences corresponding to a specific mRNA, the greater the expression of the corresponding gene.
This approach was chosen by the group of the UAB Molecular Biology of Yeast which is working in close collaboration with the company Lifesequencing. Researchers chose as a model for their study the Saccharomyces cerevisiae yeast, an organism used in basic research and in biotechnology; it was also the first eukaryote genome that was fully sequenced. A few months after a group of US scientists used this technique to build the transcriptional panorama of this organism in its resting state, the UAB research group now wishes to go a step further and define this panorama under stress conditions, by changing its pH. The project, which is currently in its final stages, also aims to validate massive sequencing techniques by comparing them to DNA microchip-based technologies, which were developed a few years ago by the same UAB research group.
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