Abstract: Hepatitis A virus (HAV) continues to cause outbreaks of viral hepatitis, which remains a socially significant disease despite existing vaccines. During the production of vaccines, HAV strains that require prolonged incubation of infected cell cultures, up to 4 weeks, are used. Several HAV strains, which are fast-growing on cell cultures, up to 7–10 days, are known. This paper presents the results of virological characterization and analysis of the genome of the HAV strain MB-7/293 (HepA-293) derived from MB-7/4647, one of the fast-growing strains, which was obtained by adapting it to HEK293 cell culture (human embryonic kidney cells). It was shown that the MB-7/293 strain significantly exceeds the parent MB-7/4647 strain in productivity when cultured on HEK293 cell culture, i.e., in the yield of the infectious virus and the virus-specific antigen. Analysis of the nucleotide sequence of full-length genomic RNA of the MB-7/293 strain showed that it differs from the genome sequence of MB-7/4647 in three nucleotide substitutions in the 5'-untranslated region and 20 nucleotide substitutions in the coding region, which lead to substitutions of ten amino acid residues in the proteins VP3, VP1, 2B and 2C. Comparative analysis of the genomic sequences of the strains MB-7/293, MB-7/4647, and other HAV strains, including the previously described fast-growing strains, allowed to reveal mutations that affect the reproductive properties of the virus.