Epstein-Barr virus (EBV) is a gamma-herpesvirus that infects epithelial cells and naïve Bcells. With a prevalence of about 95% in the world population, it is generally acquired duringchildhood or adolescence and is responsible for a non-threatening clinical condition calledinfectious mononucleosis. However, particularly in the immunosuppressed this virus isimplicated in the pathology of lymphomas and some carcinomas. The presence of EBVencodedstable intronic sequence RNAs in EBV-positive cell lines was detected for the firsttime in 2013 and their role in the EBV’s life cycle is not yet known. Here is reported thegeneration of the first virus knock-out virus for sisRNA-1 and the preliminary study of thevirus’ behaviour in B cell infection. Evidence found by monitoring cell proliferation throughmicroscopy and FACS analysis shows that knocking out the intron that encodes sisRNA-1causes proliferation impairment by day 8 after B cell infection, but doesn’t impede laterproliferation and outgrowth of LCLs. Viral transcription of LMP1 and Cp vs Wp promoterusage was analysed as well as viral protein expression of the EBNAs but no significantdifferences between WT and sis1KO where found. It remains to be determined if the effectsobserved where in fact a consequence of the lack of sisRNA-1 or due to the disruption ofBHRF1 splicing that we hypothesised to be a possible consequence of deleting sisRNA-1.
|Date of Award||10 Apr 2017|
- Universidade Católica Portuguesa
|Supervisor||Robert E. White (Supervisor)|
- Mestrado em Microbiologia Aplicada