Cameron Chambers

Evaluating the Potency of siRNA targeting Guinea Pig Cytomegalovirus

Cameron Chambers

ABSTRACT
Human cytomegalovirus (CMV) is an opportunistic herpesvirus and the most common cause of congenital viral infections. One in 200 newborns in the United States is diagnosed with congenital CMV, which can cause developmental and neurological disorders. Short interfering RNA (siRNA) are an emerging class of antiviral therapeutics. We sought to test the potency of siRNA inhibition on guinea pig (GPCMV), a virus that is used as part of a small animal model of congenital CMV. Using guinea pig lung fibroblasts and custom designed siRNA that targeted seven GPCMV genes, we investigated the inhibitory effects of siRNA on viral replication at several time points post-infection. siRNA treated cells were infected and GPCMV replication was quantified by luciferase production or plaque assay. We concluded that the most effective siRNAs targeted GP57 (major DNA-binding protein), GP86 (major capsid protein), and GP123 (IE1 transcription factor). Targeting these genes with siRNA resulted in upwards of a 100-fold reduction in GPCMV replication. Future studies will include a comprehensive characterization of these siRNAs and an in vivo trial to determine if prophylactic siRNA injection protects against congenital infection when pregnant guinea pigs are infected with GPCMV.