Analysis of VapBC TA pairs and their biotech potential

The use of toxin-antitoxin (TA) systems for biotechnological and therapeutic applications is emerging. Recent studies have revealed that the Kid and Kis TA system offers a great potential to induce selective, highly specific, and safe ablation of targeted cells in eukaryotic organisms. The major goal of this project is to extend the observations made with Kid and Kis to other TA pairs, by testing their ability to kill and/or protect eukaryotic cells in vivo and to use them to develop novel tools for the selective ablation of eukaryotic cells in vivo. VapBC pairs belong to a large family of TA pairs that are abundantly represented in bacterial and archaeal genomes. However, little is known about them. Particularly, the cellular targets, and the mode and range of action of VapC proteins remain to be described, as well as how their toxicity is neutralized by their VapB counterparts. The rationale for analyzing other TA pairs than Kid and Kis is two fold: First, different toxins have different targets, and some of these targets will be more suitable than others for certain applications. Second, increasing the number of TA pairs analyzed will contribute to the identification of determinants required for their activity and selectivity. This will lead to a better understanding of TA pairs in general and ultimately, to the rational design of novel TA pairs, particularly suited for biotechnological and therapeutic applications. To do this, we aim to characterize biochemically different members of the family of VapBC pairs, to study their mode of action, their specificities, and the effects of expressing them in prokaryotes and eukaryotes. This knowledge is essential in order to evaluate the biotechnological and therapeutic potential of VapBC pairs.