Molecular Beacon Signaling of Trichomonas vaginalis virus
Derrick L. Burt1, Peter C. Martin2, Cory G. Toyota1
1Department of Chemistry and Biochemistry, Millsaps College, Jackson, MS
2Vanderbilt University, Nashville, TN
Trichomanias is the most common sexually transmitted infection worldwide, and is caused by the protozoan parasite Trichomonas vaginalis. The parasite itself may be infected with Trichomonas vaginalis virus (TVV), a double-stranded RNA virus from the Totiviridae family. There are four known distinct subspecies of TVV, (TVV1-TVV4). Molecular beacons are oligonucleotides with a target-specific loop bookended by complementary sequences that complete the hairpin. Attached to the ends of the beacon are a fluorescence quencher on one end and a fluorophore on the other. The stability is such that in the absence of target sequences, molecular beacons remain in hairpin structure and the quencher prevents the fluorophore from emitting detectable fluorescence. In the presence of target sequence, the beacon will bind to the target and, now in linear form, begin to fluoresce upon excitation. This leads to molecular beacons’ high signal to background ratio, relative to more traditional linear probes. Another notable ability of molecular beacons is their capacity to perform at physiological temperature and conditions, allowing for real-time in vivo experimentation. We will take advantage of this feature and use molecular beacons targeted to TVV1. After first determining an in vitro affinity for TVV1 genetic sequences, we will use molecular beacons in living Trichomonas vaginalis cells, determining specificity between infected and uninfected cells. Among the infected cells, fluorescence of TVV1-bound beacons will allow for determination of real-time double-stranded RNA virus localization.