Russian Journal of Developmental Biology

, Volume 49, Issue 6, pp 356–361 | Cite as

Optimal Number of Embryos for Transplantation in Obtaining Genetic-Modified Mice and Goats

  • Yu. Yu. Silaeva
  • Yu. K. Kirikovich
  • L. N. Skuratovskaya
  • A. V. DeikinEmail author


The technology of creating genetically modified animals (placental mammals) by microinjection into the pronucleus of a fertilized egg suggests, as one of the key stages, the transplantation of early embryos into female recipients. However, there is a wide range of opinions among researchers about the optimal number of embryos to be transferred to the female recipient. Thus, data on transplantation of 20–60 mouse embryos and from 2 to 6 goat embryos to one recipient are given in the methodological literature and experimental articles devoted to the method of creating genetically modified animals. Thus, the standard recommendation is the transfer of a much larger number of embryos than that which develops in animals of both species in physiological pregnancy. At the same time, technology of transplantation of bovine embryos (cattle) involves the transfer of one embryo, which is the physiological norm for this species of animals. Clinical protocols of assisted reproductive technologies for the transplantation of human embryos also recommend the transfer of one embryo, because transferring the number of embryos greater than in physiological pregnancy leads to increased risks. In our work, we analyze the results of experiments on obtaining genetically modified mice and goats and provide data indicating the need to revise the standard recommendations on the number of transferred embryos downward. We believe that the number of transferred embryos should not exceed the number of embryos characteristic for physiological pregnancy. Excess of the number of transplanted embryos leads to a pathological course of pregnancy and a significant decrease in overall performance.


genetically modified animals mice goats embryo transplantation pregnancy pathology 



This work was performed with support from the Russian Science Foundation, project no. 16-14-00150 (2470 mouse embryos were transplanted into 269 recipients, 277 calves were obtained. The results of transplantation and statistical processing were systematized).

The work was conducted using equipment of the Center for Shared Use of the Gene Biology Institute of the Russian Academy of Sciences.

We would like to thank A.I. Budevich and I.L. Goldman for invaluable assistance in mastering the technology of creating genetically modified animals.


  1. 1.
    Amiri Yekta, A., Dalman, A. Eftekhari-Yazdi, P., et al., Production of transgenic goats expressing human coagulation factor IX in the mammary glands after nuclear transfer using transfected fetal fibroblast cells, Transgenic Res., 2013, vol. 22, no. 1, pp. 131–142.CrossRefGoogle Scholar
  2. 2.
    Baldassarre, H., Wang, B., Kafidi, N., et al., Production of transgenic goats by pronuclear microinjection of in vitro produced zygotes derived from oocytes recovered by laparoscopy, Theriogenology, 2003, vol. 59, nos. 3–4, pp. 831–839.CrossRefGoogle Scholar
  3. 3.
    Batista, R., Melo, C., Souza-Fabjan, J., Teixeira, D., et al., Phenotypic features of first-generation transgenic goats for human granulocyte-colony stimulation factor production in milk, Biotechnol. Lett., 2014, vol. 36, no. 11, pp. 2155–2162.CrossRefGoogle Scholar
  4. 4.
    Cho, A., Haruyama, N., and Kulkarni, A., Generation of Transgenic Mice, Current Protocols in Cell Biology [Internet], Hoboken, NJ, USA: John Wiley and Sons, Inc., 2009.Google Scholar
  5. 5.
    Damert, A. and Kusserow, H., Generation of transgenic mice by pronuclear injection, in Blood–Brain Barrier, New Jersey: Humana Press, 2003, pp. 513–528.Google Scholar
  6. 6.
    Deĭkin, A.V., Kovrazhkina, E.A., Ovchinnikov, R.K., et al., A mice model of amyotrophic lateral sclerosis expressing mutant human FUS protein, Zh. Nevrol. Psikhiatr. im. S.S. Korsakova, 2014, vol. 114, no. 8, pp. 62–69.Google Scholar
  7. 7.
    Deykin, A.V., Ermolkevich, T.G., Gursky, Y.G., et al., The state of health and the reproductive potential of transgenic mice secreting recombinant human lactoferrin in milk, Dokl. Biochem. Biophys., 2009, vol. 427, pp. 195–198.CrossRefGoogle Scholar
  8. 8.
    Freitas, V., Serova, I., Moura, R., et al., The establishment of two transgenic goat lines for mammary gland hG-CSF expression, Small Rumin. Res., 2012, vol. 105, nos. 1–3, pp. 105–113.CrossRefGoogle Scholar
  9. 9.
    Goldman, I., Georgieva, S., Gurskiy, Y., et al., New opportunities of using transgenic milk animals for pharmaceutical human protein production, Transgenic Res., 2012a, vol. 21, no. 4, p. 923.Google Scholar
  10. 10.
    Goldman, I., Georgieva, S., Gurskiy, Y., et al., Production of human lactoferrin in animal milk, Biochem. Cell Biol., 2012b, vol. 90, no. 3, pp. 513–519.CrossRefGoogle Scholar
  11. 11.
    Gurskiy, Y., Garbuz, D., Soshnikova, N., et al., The development of modified human Hsp70 (HSPA1A) and its production in the milk of transgenic mice, Cell Stress Chaperones, 2016, vol. 21, no. 6, pp. 1055–1064.CrossRefGoogle Scholar
  12. 12.
    Gursky, Y., Bibilashvili, R., Minashkin, M., et al., Expression of full-length human pro-urokinase in mammary glands of transgenic mice, Transgenic Res., 2009, vol. 18, no. 5, pp. 747–756.CrossRefGoogle Scholar
  13. 13.
    Hansson, L., Edlund, M., Edlund, A., et al., Expression and characterization of biologically active human extracellular superoxide dismutase in milk of transgenic mice, J. Biol. Chem., 1994, vol. 269, no. 7, pp. 5358–5363.Google Scholar
  14. 14.
    Hasler, J., Forty years of embryo transfer in cattle: a review focusing on the journal theriogenology, the growth of the industry in North America, and personal reminisces, Theriogenology, 2014, vol. 81, no. 1, pp. 152–169.CrossRefGoogle Scholar
  15. 15.
    Ittner, L. and Götz, J., Pronuclear injection for the production of transgenic mice, Nat. Protoc., 2007, vol. 2, no. 5, pp. 1206–1215.CrossRefGoogle Scholar
  16. 16.
    Kadulin, S., Ermolkevich, T., and Andreeva, L., Analysis of transfer of microinjected zygotes in production of transgenic mice, Russ. J. Dev. Biol., 2006, vol. 37, no. 2, pp. 85–89.CrossRefGoogle Scholar
  17. 17.
    Lisauskas, S., Cunha, N., Vianna, G., et al., Expression of functional recombinant human factor ix in milk of mice, Biotechnol. Lett., 2008, vol. 30, no. 12, pp. 2063–2069.CrossRefGoogle Scholar
  18. 18.
    Maksimenko, O.G., Deykin, A.V., Khodarovich, Y.M., et al., Use of transgenic animals in biotechnology: prospects and problems, Acta Naturae, 2013, vol. 5, no. 1, pp. 33–46.Google Scholar
  19. 19.
    Niavarani, A., Dehghanizadeh, S., Zeinali, S., et al., Development of transgenic mice expressing calcitonin as a beta-lactoglobulin fusion protein in mammary gland, Transgenic Res., 2005, vol. 14, no. 5, pp. 719–727.CrossRefGoogle Scholar
  20. 20.
    Pandian, Z., Marjoribanks, J., Ozturk, O., et al., Number of Embryos for Transfer Following in vitro Fertilisation or Intra-Cytoplasmic Sperm Injection, Cochrane Database of Systematic Reviews, Chichester: UK: John Wiley and Sons, Ltd., 2013.Google Scholar
  21. 21.
    Robinson, H.K., Deykin, A.V., Bronovitsky, E.V., et al., Early lethality and neuronal proteinopathy in mice expressing cytoplasm-targeted FUS that lacks the RNA recognition motif, Amyotroph. Lateral. Scler. Front. Degener., 2015, vol. 16, nos. 5–6, pp. 402–409.CrossRefGoogle Scholar
  22. 22.
    Rodriguez, A., Castro, F.O., Aguilar, A., et al., Expression of active human erythropoietin in the mammary gland of lactating transgenic mice and rabbits, Biol. Res., 1995, vol. 28, no. 2, pp. 141–153.Google Scholar
  23. 23.
    Scherzer, J., Fayrer-Hosken, R., Ray, L., et al., Advancements in large animal embryo transfer and related biotechnologies, Reprod. Domest. Anim., 2008, vol. 43, no. 3, pp. 371–376.CrossRefGoogle Scholar
  24. 24.
    Shelkovnikova, T.A., Peters, O.M., Deykin, A.V., et al., Fused in sarcoma (FUS) protein lacking nuclear localization signal (NLS) and major RNA binding motifs triggers proteinopathy and severe motor phenotype in transgenic mice, J. Biol. Chem., 2013, vol. 288, no. 35, pp. 25266–25274.CrossRefGoogle Scholar
  25. 25.
    Silaeva, Y.Y., Kalinina, A.A., Vagida, M.S., et al., Decrease in pool of T lymphocytes with surface phenotypes of effector and central memory cells under influence of TCR transgenic β-chain expression, Biochemistry (Moscow), 2013, vol. 78, no. 5, pp. 549–559.Google Scholar
  26. 26.
    Silaeva, Y.Y., Grinenko, T.S., Vagida, M.S., et al., Immune selection of tumor cells in TCR β-chain transgenic mice, J. Immunotoxicol., 2014, vol. 11, no. 4, pp. 393–399.CrossRefGoogle Scholar
  27. 27.
    Sokolov, V.E., Zhizn zhivotnykh (Life of Animals), Moscow: Prosveshchenie, 1989, vol. 7.Google Scholar
  28. 28.
    Voncken, J.W., Genetic modification of the mouse: general technology—pronuclear and blastocyst injection, in Transgenic Mouse Methods and Protocols, Totowa, NJ: Humana Press, 2011, pp. 11–36.Google Scholar
  29. 29.
    Yu, H., Chen, J., Sun, W., et al., The dominant expression of functional human lactoferrin in transgenic cloned goats using a hybrid lactoferrin expression construct, J. Biotechnol., 2012, vol. 161, no. 3, pp. 198–205.CrossRefGoogle Scholar
  30. 30.
    Yu, H., Chen, J., Liu, S., et al., Large-scale production of functional human lysozyme in transgenic cloned goats, J. Biotechnol., 2013, vol. 168, no. 4, pp. 676–683.CrossRefGoogle Scholar
  31. 31.
    Zander-Fox, D.L., Tremellen, K., and Lane, M., Single blastocyst embryo transfer maintains comparable pregnancy rates to double cleavage-stage embryo transfer but results in healthier pregnancy outcomes: the benefits of single blastocyst transfer, Aust. N. Z. J. Obstet. Gynaecol., 2011, vol. 51, no. 5, pp. 406–410.CrossRefGoogle Scholar
  32. 32.
    Zhang, J., Li, L., Cai, Y., et al., Expression of active recombinant human lactoferrin in the milk of transgenic goats, Protein Expr. Purif., 2008, vol. 57, no. 2, pp. 127–135.CrossRefGoogle Scholar
  33. 33.
    Zvezdova, E.S., Silaeva, Y.Y., Vagida, M.S., et al., Generation of transgenic animals expressing the α and β chains of the autoreactive t-cell receptor, Mol. Biol., 2010, vol. 44, no. 2, p. 277.Google Scholar

Copyright information

© Pleiades Publishing, Inc. 2018

Authors and Affiliations

  • Yu. Yu. Silaeva
    • 1
  • Yu. K. Kirikovich
    • 3
  • L. N. Skuratovskaya
    • 2
  • A. V. Deikin
    • 1
    • 2
    Email author
  1. 1.Gene Biology Institute, Russian Academy of SciencesMoscowRussia
  2. 2.Research Institute of General Pathology and PathophysiologyMoscowRussia
  3. 3.Scientific and Practical Center for Animal BreedingZhodinoRepublic of Belarus

Personalised recommendations