Analysis of the Expression and Function of the EBV-Encoded Small RNAs, the EBERs, in Heterologous Cells

The two small virally encoded RNA species, EBER-1 and EBER-2, are abundantly expressed in almost all Epstein-Barr virus (EBV)-infected cell types. Their functions in relation to the physiology of the virus remain enigmatic. In recent years, the main interest in the EBERs has been in connection with the use of these RNAs as targets for identification of EBV infection using in situ hybridization. However evidence for a possible function for the EBERs is now emerging and it seems likely that these small RNAs constitute another weapon in the armory used by EBV to infect and immortalize the host cell. EBER-1 and EBER-2 are uncapped, nonpolyadenylated, untranslated RNAs of 167 and 172 nucleotides, respectively. Even in cells with the most restricted range of EBV gene expression (e.g., most Burkitt’s lymphomas), where EBNA-1 is the only viral protein synthesized, the EBERs are always present. The two RNAs are also found in cells in which EBV replication is actively occurring. In this chapter we describe techniques for the expression of the EBERs in EBV-negative cells, and for the analysis of the functions of these small RNAs in the control of cellular protein synthesis.