{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,24]],"date-time":"2026-01-24T05:32:05Z","timestamp":1769232725162,"version":"3.49.0"},"reference-count":26,"publisher":"Institute of Cytology and Genetics, SB RAS","issue":"4","license":[{"start":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T00:00:00Z","timestamp":1657152000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/vavilov.elpub.ru\/jour\/about\/editorialPolicies#openAccessPolicy"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Vestn. VOGiS"],"abstract":"<jats:p>Due to cessation of mass smallpox vaccination in 1980, the collective immunity of humans against orthopoxvirus infections has virtually been lost. Therefore, the risk of spreading zoonotic human orthopoxvirus infections caused by monkeypox and cowpox viruses has increased in the world. First-generation smallpox vaccines based on Vaccinia virus (VAC) are reactogenic and therefore not suitable for mass vaccination under current conditions. This necessitates the development of modern safe live vaccines based on VAC using genetic engineering. We created the VAC\u03946 strain by transient dominant selection. In the VAC\u03946 genome, five virulence genes were intentionally deleted, and one gene was inactivated by inserting a synthetic DNA fragment. The virus was passaged 71 times in CV-1 cells to obtain the VAC\u03946 strain from the VAC LIVP clonal variant. Such a long passage history might have led to additional off-target mutations in VAC\u03946 compared to the original LIVP variant. To prevent this, we performed a genome-wide sequencing of VAC LIVP, VAC\u03946, and five intermediate viral strains to assess possible off-target mutations. A comparative analysis of complete viral genomes showed that, in addition to target mutations, only two nucleotide substitutions occurred spontaneously when obtaining VAC\u03944 from the VAC\u03943 strain; the mutations persisting in the VAC\u03945 and VAC\u03946 genomes. Both nucleotide substitutions are located in intergenic regions (positions 1431 and 189738 relative to LIVP), which indicates an extremely rare occurrence of off-target mutations when using transient dominant selection to obtain recombinant VAC variants with multiple insertions\/deletions. To assess the genome stability of the resulting attenuated vaccine strain, 15 consecutive cycles of cultivation of the industrial VAC\u03946 strain were performed in 4647 cells certified for vaccine production in accordance with the \u201cGuidelines for Clinical Trials of Medicinal Products\u201d. PCR and sequencing analysis of six DNA fragments corresponding to the regions of disrupted genes in VAC\u03946 showed that all viral DNA sequences remained unchanged after 15 passages in 4647 cells.<\/jats:p>","DOI":"10.18699\/vjgb-22-48","type":"journal-article","created":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T07:25:55Z","timestamp":1657178755000},"page":"394-401","source":"Crossref","is-referenced-by-count":3,"title":["Genome stability of the vaccine strain VAC\u22066"],"prefix":"10.18699","volume":"26","author":[{"given":"R. A.","family":"Maksyutov","sequence":"first","affiliation":[{"name":"State Research Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0496-390X","authenticated-orcid":true,"given":"S. N.","family":"Yakubitskiy","sequence":"additional","affiliation":[{"name":"State Research Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"I. V.","family":"Kolosova","sequence":"additional","affiliation":[{"name":"State Research Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T. V.","family":"Tregubchak","sequence":"additional","affiliation":[{"name":"State Research Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"A. N.","family":"Shvalov","sequence":"additional","affiliation":[{"name":"State Research Center of Virology and Biotechnology \u201cVector\u201d, Rospotrebnadzor"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E. 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Modified vaccinia virus Ankara undergoes limited replication in human cells and lacks several immunomodulatory proteins: implications for use as a human vaccine. J. Gen. Virol. 1998;79(Pt. 5):1159-1167. DOI 10.1099\/0022-1317-79-5-1159.","DOI":"10.1099\/0022-1317-79-5-1159"},{"key":"ref3","doi-asserted-by":"crossref","unstructured":"Drexler I., Heller K., Wahren B., Erfle V., Sutter G. Highly attenuated modified vaccinia virus Ankara replicates in baby hamster kidney cells, a potential host for virus propagation, but not in various human transformed and primary cells. J. Gen. Virol. 1998;79(Pt. 2): 347-352. DOI 10.1099\/0022-1317-79-2-347.","DOI":"10.1099\/0022-1317-79-2-347"},{"key":"ref4","doi-asserted-by":"crossref","unstructured":"Eto A., Saito T., Yokote H., Kurane I., Kanatani Y. Recent advances in the study of live attenuated cell-cultured smallpox vaccine LC16m8. Vaccine. 2015;33(45):6106-6111. DOI 10.1016\/j.vaccine.2015.07.111.","DOI":"10.1016\/j.vaccine.2015.07.111"},{"key":"ref5","doi-asserted-by":"crossref","unstructured":"Falkner F.G., Moss B. Transient dominant selection of recombinant vaccinia viruses. J. Virol. 1990;64(6):3108-3111. DOI 10.1128\/JVI.64.6.3108-3111.1990.","DOI":"10.1128\/jvi.64.6.3108-3111.1990"},{"key":"ref6","unstructured":"Guidelines for Clinical Trials of Medicinal Products (Immunobiological Medicinal Products). Part 2. Moscow: Grif and K Publ., 2012. (in Russian)"},{"key":"ref7","doi-asserted-by":"crossref","unstructured":"Katoh K., Misawa K., Kuma K., Miyata T. MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res. 2002;30(14):3059-3066. DOI 10.1093\/nar\/gkf436.","DOI":"10.1093\/nar\/gkf436"},{"key":"ref8","doi-asserted-by":"crossref","unstructured":"Kidokoro M., Shida H. Vaccinia virus LC16m8\u2206 as a vaccine vector for clinical applications. Vaccines. 2014;2(4):755-771. DOI 10.3390\/vaccines2040755.","DOI":"10.3390\/vaccines2040755"},{"key":"ref9","doi-asserted-by":"crossref","unstructured":"Kretzschmar M., Wallinga J., Teunis P., Xing S., Mikolajczyk R. Frequency of adverse events after vaccination with different vaccinia strains. PLoS Med. 2006;3(8):e272. DOI 10.1371\/journal.pmed.0030272.","DOI":"10.1371\/journal.pmed.0030272"},{"key":"ref10","doi-asserted-by":"crossref","unstructured":"Li H., Durbin R. Fast and accurate short read alignment with Burrows\u2013 Wheeler transform. Bioinformatics. 2009;25(14):1754-1760. DOI 10.1093\/bioinformatics\/btp324.","DOI":"10.1093\/bioinformatics\/btp324"},{"key":"ref11","doi-asserted-by":"crossref","unstructured":"Li H., Handsaker B., Wysoker A., Fennell T., Ruan J., Homer N., Marth G., Abecasis G., Durbin R. The sequence alignment\/map format and SAMtools. Bioinformatics. 2009;25(16):2078-2079. DOI 10.1093\/bioinformatics\/btp352.","DOI":"10.1093\/bioinformatics\/btp352"},{"key":"ref12","doi-asserted-by":"crossref","unstructured":"McKenna A., Hanna M., Banks E., Sivachenko A., Cibulskis K., Kernytsky A., Garimella K., Altshuler D., Gabriel S., Daly M., DePristo M.A. The genome analysis Toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res. 2010;20(9):1297-1303. DOI 10.1101\/gr.107524.110.","DOI":"10.1101\/gr.107524.110"},{"key":"ref13","doi-asserted-by":"crossref","unstructured":"Moss B. Smallpox vaccines: Targets of protective immunity. Immunol. Rev. 2011;239(1):8-26. DOI 10.1111\/j.1600-065X.2010.00975.x.","DOI":"10.1111\/j.1600-065X.2010.00975.x"},{"key":"ref14","doi-asserted-by":"crossref","unstructured":"Nolen L.D., Osadebe L., Katomba J., Likofata J., Mukadi D., Monroe B., Doty J., Hughes C.M., Kabamba J., Malekani J., Bomponda P.L., Lokota J.I., Balilo M.P., Likafi T., Lushima R.S., Ilunga B.K., Nkawa F., Pukuta E., Karhemere S., Tamfum J.J., Nguete B., Wemakoy E.O., McCollum A.M., Reynolds M.G. Extended human-to-human transmission during a monkeypox outbreak in the Democratic Republic of the Congo. Emerg. Infect. Dis. 2016;22(6):1014-1021. DOI 10.3201\/eid2206.150579.","DOI":"10.3201\/eid2206.150579"},{"key":"ref15","doi-asserted-by":"crossref","unstructured":"Okonechnikov K., Golosova O., Fursov M., UGENE team. Unipro UGENE: a unified bioinformatics toolkit. Bioinformatics. 2012; 28(8):1166-1167. DOI 10.1093\/bioinformatics\/bts091.","DOI":"10.1093\/bioinformatics\/bts091"},{"key":"ref16","doi-asserted-by":"crossref","unstructured":"Reynolds M.G., Doty J.B., McCollum A.M., Olson V.A., Nakazawa Y. Monkeypox re-emergence in Africa: a call to expand the concept and practice of One Health. Expert Rev. Anti Infect. Ther. 2019;17(2): 129-139. DOI 10.1080\/14787210.2019.1567330.","DOI":"10.1080\/14787210.2019.1567330"},{"key":"ref17","doi-asserted-by":"crossref","unstructured":"Robinson J.T., Thorvaldsdottir H., Winckler W., Guttman M., Lander E.S., Getz G., Mesirov J.P. Integrative genomics viewer. Nat. Biotechnol. 2011;29(1):24-26. DOI 10.1038\/nbt.1754.","DOI":"10.1038\/nbt.1754"},{"key":"ref18","doi-asserted-by":"crossref","unstructured":"Sanchez-Sampedro L., Perdiguero B., Mejias-Perez E., Garcia-Arriaza J., Di Pilato M., Esteban M. The evolution of poxvirus vaccines. Viruses. 2015;7(4):1726-1803. DOI 10.3390\/v7041726.","DOI":"10.3390\/v7041726"},{"key":"ref19","doi-asserted-by":"crossref","unstructured":"Shchelkunov S.N. Emergence and reemergence of smallpox: the need in development of a new generation smallpox vaccine. Vaccine. 2011;29(Suppl. 4):D49-D53. DOI 10.1016\/j.vaccine.2011.05.037.","DOI":"10.1016\/j.vaccine.2011.05.037"},{"key":"ref20","doi-asserted-by":"crossref","unstructured":"Shchelkunov S.N. An increasing danger of zoonotic orthopoxvirus infections. PLoS Pathog. 2013;9(12):e1003756. DOI 10.1371\/journal.ppat.1003756.","DOI":"10.1371\/journal.ppat.1003756"},{"key":"ref21","doi-asserted-by":"crossref","unstructured":"Shchelkunov S.N., Shchelkunova G.A. Genes that control vaccinia virus immunogenicity. Acta Naturae. 2020;12(1):33-41. DOI 10.32607\/actanaturae.10935.","DOI":"10.32607\/actanaturae.10935"},{"key":"ref22","doi-asserted-by":"crossref","unstructured":"Singh R.K., Balamurugan V., Bhanuprakash V., Venkatesan G., Hosamani M. Emergence and reemergence of vaccinia-like viruses: global scenario and perspectives. Indian J. Virol. 2012;23(1):1-11. DOI 10.1007\/s13337-012-0068-1.","DOI":"10.1007\/s13337-012-0068-1"},{"key":"ref23","unstructured":"Smallpox and its Eradication. Geneva: World Health Organization, 1988."},{"key":"ref24","doi-asserted-by":"crossref","unstructured":"Volz A., Sutter G. Modified vaccinia virus Ankara. History, value in basic research, and current perspectives for vaccine development. Adv. Virus Res. 2017;97:187-243. DOI 10.1016\/bs.aivir.2016.07.001.","DOI":"10.1016\/bs.aivir.2016.07.001"},{"key":"ref25","doi-asserted-by":"crossref","unstructured":"Yakubitskiy S.N., Kolosova I.V., Maksyutov R.A., Shchelkunov S.N. Attenuation of vaccinia virus. Acta Naturae. 2015;7(4):113-121. DOI 10.32607\/20758251-2015-7-4-113-121.","DOI":"10.32607\/20758251-2015-7-4-113-121"},{"key":"ref26","doi-asserted-by":"crossref","unstructured":"Yakubitskiy S.N., Kolosova I.V., Maksyutov R.A., Shchelkunov S.N. Highly immunogenic variant of attenuated vaccinia virus. Dokl. Biochem. Biophys. 2016;466:35-38. 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