Rolosense: An innovative platform for automatic mobile phone readout of active SARS-CoV-2 particles

Project AbstractThe ultimate goal of this proposal is to develop a novel platform technology for automatic surveillanceand tracing of airborne SARS-CoV-2 virus particles in real time. The centerpiece of this proposal is the"Rolosense" technology which leverages a DNA micromotor as the virus sensing and transductionmaterial (VSTM) that can be detected by a conventional smart phone camera. This provides bothgeographical tracing and surveillance. Rolosense motor are comprised a DNA-coated spherical particle(5 micrometer diameter) that hybridizes to a surface modified with complementary RNA. The particlemoves at speeds of over 1 micron/minute upon the addition of RNase H, which selectively hydrolyseshybridized RNA but not single-stranded RNA. DNA motors coated with virus binding ligand (VBL) stallin presence of SARS-CoV-2 virus particles. Because motors move autonomously for distances up tomillimeters without intervention, the assay is fully automated, and conventional steps such as viralinactivation, RNA isolation and amplification are not required. The readout is performed using anautomated smart phone app for particle tracking without the need for a spectrophotometer orfluorometer. Preliminary data shows realtime SARS-CoV-2 pseudovirus particle sensing. Milestonesinclude the screening and identification of high affinity and high specificity VBLs. Both aptamers andantibody VBLs will be screening and validated. Simulations and experiments will be used to understandthe role of temperature and environmental conditions in modulating Rolosense performance.Multivalent display of VBLs with DNA origami will enhance avidity. Finally, microfluidic chips withairborne droplet capture will be implemented and tested. The work will be performed by a highlyinterdisciplinary team with complementary expertise and a track-record of co-publications. PI Salaitainvented the Rolosense technology and has past experience in developing cell phone diagnostics andsynthetic motors. Co-I Melikian is an expert virologist, Co-I Heemstra is an expert at developingaptamers for novel targets, Co-I Ke has extensive experience in DNA origami structures for avid targetbinding and has co-authored work on Rolosense, Co-I Rajaraman and Primordia are experts atmicrofluidic device development and commercialization. Our solution offers the potential to provide animmediate solution to today's urgent virus sensing and tracing needs.