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Coronavirus reverse genetics has greatly contributed to our current understanding of coronavirus molecular biology and diseases. Starting in the early 1990's reverse genetic techniques have been applied to the modification of the 3'-third of coronavirus genomes by targeted recombination. In 2000, reverse genetic systems based on full-length cDNAs made the entire coronavirus genome amenable to mutagenesis. Coronavirus reverse genetic techniques have been applied to the analysis of all aspects of coronavirus infections. These include studies on the tropism, pathogenesis, virus entry, virion formation and RNA synthesis of coronaviruses. In addition to the generation of recombinant coronaviruses, reverse genetic approaches have also been used for the development of coronavirus replicon RNAs and novel vectors designed for heterologous gene expression.