Kyasanur Forest Disease Virus (KFDV) is a tick-borne, hemorrhagic fever-causing member of the Flaviviridae. With infections annually ranging from 50 to 1000 people in south-west India and the lack of effective treatments, a better understanding of this virus is needed. The development of a Reverse Genetics System for KFDV would provide the opportunity to address these issues in future studies. Using molecular techniques, the KFDV genome sequence was elucidated and the reverse genetics system was created. Utilizing this system live, infectious KFDV particles were produced from mammalian cell culture, thereby validating the success of the reverse genetics system. The implementation of this system will enable researchers to better study pathogenesis and disease progression, virus-host interplay, virion structure, genome replication and the emergence of effective therapeutics and vaccines.