ÓÄÊ 619:636.082:606:636.03
DOI 10.33861/2071-8020-2023-4-9-13
Donnik I. M., Makutina V. A., Isayeva A. G., Krivonogova A. S., Mymrin V. S.
Summary. The problematic issues of modern dairy cattle breeding and replenishment of breeding resources are presented. The key problems of the industry are outlined and analyzed: dependence on import breeding material, inbreeding, the use of outdated methods of assessing the breeding value of animals, reduction of the animal longevity, deterioration of the health of highly productive livestock, the disappearance of domestic cattle breeds. It has been shown that modern genetic technologies allow to not only preserve the biodiversity of cattle breeds and reproduce the most valuable animals, but also make it possible to speed up the breeding process. The CRISPR/Cas9-based bovine genome editing technology made it possible to obtain preimplantation embryos with knockout genes encoding the CD209 receptor and the milk allergenic protein betalactoglobulin (BLG). An improved and species-specific adapted bovine cell processing technique from donor biomaterial collection to cryopreservation of mature oocytes and edited embryos achieved a fairly high level of zygote survival after editing. Depending on the method of delivery of the editing system, the knockout rate ranged from 14% to 30%. The most effective methods of CRISPR/Cas9 delivery to the zygote were oocyte transduction by adeno-associated virus AAV2 and microinjection of Cas9 RNA and sgRNA against the BLG and CD209 genes into the zygote cytoplasm. Gene editing techniques are promising for obtaining bovine embryos suitable for transplantation with the desired economically significant traits. The use of bovine gene editing technology makes it possible to narrowly target changes in the genome regions associated with individual traits while generally preserving the genetic potential of the most valuable breeding farm animals.
Keywords: livestock breeding, selection, gene editing, bovine embryos, CRISPR/Cas9, beta-lactoglobulin, CD209, gene editing system delivery, adeno-associated virus, RNA microinjection.
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Author affiliation:
Donnik Irina M., D. Sc. in Biology, professor, Academician of the Russian Academy of Sciences, chief scientific researcher of the department of monitoring and forecasting of infectious diseases of the Ural Federal Agrarian Scientific Research Centre of the Ural Branch of the Russian Academy of Sciences; 112-a, Belinskogo st., Ekaterinburg, 620145; e-mail: ktqrjp7@yandex.ru.
Makutina Valeriya A., Ph. D. in Biology, senior scientific researcher of the laboratory of genetic research and animal selection of the Ural Federal Agrarian Scientific Research Centre of the Ural Branch of the Russian Academy of Sciences; 112-a, Belinskogo st., Ekaterinburg, 620145; e-mail: makutina_v@rambler.ru.
Krivonogova Anna S., D. Sc. in Biology, leading scientific researcher of the laboratory of biological technology of the Ural Federal Agrarian Scientific Research Centre of the Ural Branch of the Russian Academy of Sciences; 112-a, Belinskogo st., Ekaterinburg, 620145; phone: 8-343-2572044; e-mail: tel-89826512934@yandex.ru.
Mymrin Vladimir S., D. Sc. in Biology, senior scientific researcher of the laboratory of genetic research and animal selection of the Ural Federal Agrarian Scientific Research Centre of the Ural Branch of the Russian Academy of Sciences; 112-a, Belinskogo st., Ekaterinburg, 620145; e-mail: mimrin@mail.ru.
Responsible for correspondence with the editorial board: Isaeva Albina G., D. Sc. in Biology, leading scientific researcher of laboratory of biological technology of the Ural Federal Agrarian Scientific Research Centre of the Ural Branch of the Russian Academy of Sciences; 112-a, Belinskogo st., Ekaterinburg, 620145; phone: 8-343-2572044; e-mail: isaeva.05@bk.ru.
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