Rabies is a disease of the central nervous system caused by the Rabies virus. Current methods to test for Rabies are costly, time consuming, and not sensitive enough to identify the disease early enough for treatment. A novel mechanism of virus identification that has shown promise for Ebola and Zika virus detection is the toehold switch. Toehold switches are RNA-based constructs that allow the visual identification of genomic targets of interest. They are composed of a switch RNA and a trigger RNA. In the presence of the trigger, a change in secondary structure causes the reporter gene following the switch to be expressed. When the trigger is not
present, the gene is repressed. This project designed a toehold switch for use as a Rabies virus sensor. First, a region of the Rabies genome was identified as a promising trigger sequence. From this sequence, a complementary switch was designed with GFP as a reporter gene. The designed switch was constructed and inserted into a pBluescript plasmid backbone. The next steps of this project would be to develop an assay to test the effectiveness of the switch to identify Rabies virus.
