5-Azacytidine treatment of Syrian hamster (Mesocricetus auratus) cells, BHK21-cl.13, and primary embryo fibroblasts, activates the production of intracisternal type R particles (IRP) as ascertained by electron microscopy scanning and counting. Efficiency of the activation is dose- and time of treatment-dependent. The transcription of mouse intracisternal A particle (IAP) related sequences, which have been described in Syrian hamster genomic DNA, is increased at the same time. This correlates with demethylation of hamster IAP related genes, as shown by restriction of high molecular weight DNA from 5-azacytidine treated cells with methylation sensitive enzymes, and subsequent electrophoresis and molecular hybridization of the fragments with specific DNA probes. In 5-azacytidine treated cells derived from other hamster species (Cricetulus griseus and C. migratorius), intracisternal R particles were not induced, nor were mouse IAP-related RNAs detected.