Minigenome System to Study Respiratory Syncytial Virus Transcription Versus Genome Replication.

The respiratory syncytial virus (RSV) minigenome system is a powerful analytical tool that has enabled viral transcription and genome replication to be modeled in a cellular context without viral infection. The minigenome assay can be used to dissect the different steps in RSV transcription and genome replication, assess the effects of mutations in cis-acting RNA elements, analyze mutations in the polymerase and other proteins in the transcription-replication machinery, and assess the effect of antiviral compounds on RSV polymerase functions. Here, we describe the RSV minigenome assay and detail a northern blotting technique for detecting viral mRNA transcripts and genome replication products.