Download Isolation of HSV-1 Small RNA Interference Molecules in Latently

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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.