Project description DEENESFRITPL Adaptive immunity in bacteria To survive in a hostile environment under constant attacks from viruses, microbes have evolved an adaptive antiviral system called CRISPR-Cas. The system incorporates invader DNA sequences in a memory locus (CRISPR), and forms guide RNAs from this locus, followed by the degradation of invading target DNA using CRISPR RNA-guided protein complexes. The point mutations in the targeted region of the invader’s DNA are recognised by the system by integrating new memory sequences against the same invader via a priming process. The ERC-funded REMEMBER project aims to uncover the mechanism of memory formation in CRISPR against mutated viruses that infect bacteria. The study will test the hypothesis that perfectly matching and degenerate targets trigger simultaneous virus DNA degradation and memory updates. Show the project objective Hide the project objective Objective Microbes in natural ecosystems are under constant evolutionary pressure from viruses. To survive in this hostile environment microbes have evolved an adaptive immune system called CRISPR-Cas. The immune system is based on incorporation of invader DNA sequences in a memory locus (CRISPR), the formation of guide RNAs from this locus, and the degradation of invading target DNA using CRISPR RNA-guided protein complexes. Invaders escape immunity by making point mutations in the targeted region of their DNA, but hosts quickly restore immunity by integrating new memory sequences against the same invader in a process called priming. Recently, I have made the remarkable discovery that hosts mount a primed immune response even when facing heavily mutated invaders. This implies that the memory of the CRISPR-Cas system not only functions in the short term against relatively recent threats, but also remembers a range of revisiting old foes in the long term, providing a huge evolutionary benefit for the host in the arms race with their invaders. This proposal sets out to determine the mechanism of the enigmatic process of primed memory formation against heavily mutated invaders. Using a combination of genetic, biochemical and structural approaches, including state-of-the-art single molecule imaging of CRISPR immunity in living Escherichia coli cells, I will investigate the driving hypothesis that perfectly matching and degenerate targets are differentially recognized, and trigger either target DNA degradation or priming. Moreover, I will test the supposition that degenerate priming is a universal phenomenon among different CRISPR-Cas types. If this is the case, degenerate priming will impair the use of viruses as therapeutic agents to treat antibiotic resistant bacterial infections. To prevent CRISPR resistance I propose to screen for organic molecules that inhibit the formation of CRISPR resistance. These molecules can be co-administered with viruses to potentiate treatments. Fields of science medical and health sciencesmedical biotechnologygenetic engineeringgene therapynatural sciencesbiological sciencesmicrobiologybacteriologynatural sciencesbiological sciencesmicrobiologyvirologynatural sciencesbiological sciencesgeneticsDNAmedical and health sciencesbasic medicinepharmacology and pharmacydrug resistanceantibiotic resistance Programme(s) H2020-EU.1.1. - EXCELLENT SCIENCE - European Research Council (ERC) Main Programme Topic(s) ERC-StG-2014 - ERC Starting Grant Call for proposal ERC-2014-STG See other projects for this call Funding Scheme ERC-STG - Starting Grant Host institution TECHNISCHE UNIVERSITEIT DELFT Net EU contribution € 1 373 296,84 Address STEVINWEG 1 2628 CN Delft Netherlands See on map Region West-Nederland Zuid-Holland Delft en Westland Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 1 373 296,84 Beneficiaries (2) Sort alphabetically Sort by Net EU contribution Expand all Collapse all TECHNISCHE UNIVERSITEIT DELFT Netherlands Net EU contribution € 1 373 296,84 Address STEVINWEG 1 2628 CN Delft See on map Region West-Nederland Zuid-Holland Delft en Westland Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 1 373 296,84 WAGENINGEN UNIVERSITY Participation ended Netherlands Net EU contribution € 125 886,91 Address DROEVENDAALSESTEEG 4 6708 PB Wageningen See on map Region Oost-Nederland Gelderland Veluwe Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Total cost € 125 886,91
