Abstract
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.
Similar content being viewed by others
REFERENCES
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.
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.
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.
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.
Damert, A. and Kusserow, H., Generation of transgenic mice by pronuclear injection, in Blood–Brain Barrier, New Jersey: Humana Press, 2003, pp. 513–528.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Ittner, L. and Götz, J., Pronuclear injection for the production of transgenic mice, Nat. Protoc., 2007, vol. 2, no. 5, pp. 1206–1215.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Sokolov, V.E., Zhizn zhivotnykh (Life of Animals), Moscow: Prosveshchenie, 1989, vol. 7.
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.
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.
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.
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.
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.
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.
ACKNOWLEDGMENTS
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.
Author information
Authors and Affiliations
Corresponding author
Additional information
Translated by P. Kuchina
Rights and permissions
About this article
Cite this article
Silaeva, Y.Y., Kirikovich, Y.K., Skuratovskaya, L.N. et al. Optimal Number of Embryos for Transplantation in Obtaining Genetic-Modified Mice and Goats. Russ J Dev Biol 49, 356–361 (2018). https://doi.org/10.1134/S106236041806005X
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1134/S106236041806005X