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45833 Isolation of HSV-1 Small RNA Interference Molecules in Latently Infected Rabbit TG Presenter: Teresa C Lee Mentor: Guey-Chuen Perng The herpes simplex virus type I (HSV-1) becomes dormant once infecting its host in sensory neurons of the trigeminal ganglia (TG). However, how HSV-1 establishes life-long latency is yet to be understood. Studies have shown all organisms have some type of counteractive mechanism towards their host’s immune system. Small RNA interference molecule (RNAi) is one such mechanism. RNAi, a homology-based silencing system, introduces double-stranded RNA into a cell, resulting in targeted silencing of gene expression. During HSV neuronal latency, the major gene actively transcribing is the latency associated transcript (LAT) gene. Thus, LAT is the distinct feature of HSV neuronal latency. RNAi is expected to be present in the trigeminal ganglia (TG) of HSV latently infected animals. By isolating and studying RNAi, we may gain better understanding of how HSV-1 remains latent in humans by studying immune competent hosts such as mice. We first tested our theory in HSV infected culture and total RNA was extracted from cultured rabbit skin (RS) and monkey kidney (CV-1) cells. The extracted RNA was separated by running them on 3% agaroseformaldehyde gel electrophoresis and transferred onto a nylon membrane via Northern capillary method. The membrane was dried, UV-crosslinked, and hybridized by a P-32 isotope HSV LAT DNA fragment. Probed RNA could then be visualized by autoradiography. Results showed the presence of small RNA molecules in infected cells and this suggested that isolation of RNAi was feasible in latently infected neurons.