PhD defence

The Journey of Prokaryotic Argonaute Proteins. From fundamentals to application


Argonaute proteins (Agos) form a diverse group of enzymes found in every domain of life. The Journey of prokaryotic Argonaute proteins (pAgos) described in this thesis begins with the characterization of a newly discovered pAgo from Clostridium butyricum (CbAgo). This Ago is the first example of a pAgo that efficiently cleaves single stranded DNA at 37°C, thereby paving the road towards the development of pAgo-based applications at moderate temperature, such as genome editing. The Journey ends with the successful utilization of previously characterized thermophilic pAgos from Thermus thermophilus (TtAgo) and Pyrococcus furiosus (PfAgo) in two novel pAgo-based molecular applications.