Direct access Research units
Back

Roles of non-coding RNAs in a physiologically relevant viral infection

Mouse 3T3 fibroblasts
Mouse 3T3 fibroblasts

In addition to EBV and KSHV, we also analyze extensively the small RNAs of cells infected with the betaherpesvirus mouse cytomegalovirus (MCMV). This exhaustive sequencing analysis will eventually lead to the determination of the prognosis power of small RNA profiling. The small RNA analysis has also been refined by looking at their association with different members of the Argonaute family. We are working toward determining the mechanisms responsible for the variations in expression of both cellular and viral miRNAs. The expression of small RNAs can be modulated either at a transcriptional level, or posttranscriptionally at the processing or stability level.

Small RNA cloning protocol
Small RNA cloning protocol

In order to get insight into the real importance of viral small RNAs in a physiologically relevant infection, we use mutant clones of MCMV in which we have deleted individual miRNA genes and we measure their infectiosity in vitro and in vivo.We also make use of a mouse model of Dicer to measure the global importance of miRNAs in the MCMV infection. We have access to both a hypomorphic Dicer mutant, and to an inducible floxed Dicer strain. We aim at determining the importance of Dicer in acute infection and in the reactivation of latent MCMV infection.

MCMV in vivo
Detection of MCMV miRNAs by nuclease protection assay (A) and determination of viral titers (B) in different mouse organs