dsRNA-based detection of viruses – VDOSAGE

Double-stranded RNA (dsRNA) is a hallmark of a vast majority of viral infections, it is produced by RNA viruses during replication and by DNA viruses upon convergent transcription. Despite being largely unique to viruses, dsRNA-centered approaches for virus detection and surveillance are largely underexplored. The principal aim of VDOSAGE is the proof-of-concept that dsRNA monitoring is an easy, quantitative and specific means for virus surveillance in living organisms, including arthropods and plants. By exploiting the remarkable dsRNA-binding properties of B2 protein from Flock house virus and to-be-developed Nanobodies (Nbs) with high affinity for dsRNA, we propose to build a state-of-the-art, portable device for ultrasensitive quantification of dsRNA using surface plasmon resonance (dsSPR). In parallel, we aim to develop dsNanoSeq by combining dsRNA-capture with direct RNA sequencing using Oxford nanopore technology to allow real time identification of viruses. Once optimized under laboratory conditions, dsSPR and dsNanoSeq will be validated with field-grown plants, field-caught mosquitoes and human plasma cells. To meet these objectives, we will leverage the unique know-how and expertise of four partners: B2/Nbs, dsRNA technology and plant virology (Partner 1, P1), animal virology (P2), analytical chemistry and biotechnology (P3), direct RNA sequencing (P4). The project combines innovative, multidisciplinary approaches to address future eco-epidemiological risks. In addition, VDOSAGE will improve our understanding of plant and animal viromes, as well as virus epitranscriptomic marks. Centered on the One Health perspective, VDOSAGE will improve virus surveillance in humans, animals and plants.