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Journal of equine veterinary science2021; 99; 103394; doi: 10.1016/j.jevs.2021.103394

Genetic Manipulation of the Equine Oocyte and Embryo.

Abstract: As standard in vitro fertilization is not a viable technique in horses yet, many different techniques have been used to create equine embryos for research purposes. One such method is parthenogenesis in which an oocyte is induced to mature into an embryo-like state without the introduction of a spermatozoon, and thus they are not considered true embryos. Another method is somatic cell nuclear transfer (SCNT), in which a somatic cell nucleus from an extant horse is inserted into an enucleated oocyte, creating a genetic clone of the donor horse. Due to limited availability of equine oocytes in the United States, researchers have investigated the potential for combining equine somatic cell nuclei with oocytes from other species to make embryos for research purposes, which has not been successful to date. There has also been a rising interest in producing transgenic animals using sperm exposed to exogenous DNA. The successful creation of transgenic equine blastocysts shows the promise of sperm mediated gene transfer (SMGT), but this method is not ideal for other applications, like gene therapy, because it cannot be used to induce targeted mutations. That is why technologies like CRISPR/Cas9 are vital. In this review, we argue that parthenogenesis, SCNT, and interspecies SCNT can be considered genetic manipulation strategies as they create embryos that are genetically identical to their parent cell. Here, we describe how these methods are performed and their applications and we also describe the few methods that have been used to directly modify equine embryos: SMGT and CRISPR/Cas9.
Copyright © 2021 Elsevier Inc. All rights reserved.
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Publication Date: 2021-02-03 PubMed ID: 33781418PubMed Central: PMC8605602DOI: 10.1016/j.jevs.2021.103394Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article provides an overview of various techniques used to genetically manipulate horse oocytes and embryos for research purposes as standard in vitro fertilization isn’t an option for this species. It explores different approaches including parthenogenesis, somatic cell nuclear transfer (SCNT), interspecies SCNT, and the creation of transgenic animals through sperm mediated gene transfer (SMGT) and CRISPR/Cas9 technologies.

Parthenogenesis, SCNT, and interspecies SCNT

  • Parthenogenesis is a process used to induce an oocyte, or an immature egg cell, to mature into an embryo-like state without the involvement of a sperm cell. Such entities aren’t true embryos, though, because they lack genetic material from a father.
  • Somatic cell nuclear transfer (SCNT) is another method described in the paper. It involves transferring the nucleus from a somatic cell of a horse into an oocyte that has had its nucleus removed. This process produces a genetic clone of the horse from which the somatic cell was taken.
  • Due to the limited supply of equine oocytes in the U.S, researchers have investigated the viability of interspecies SCNT – combining horse somatic cell nuclei with oocytes from different species to produce embryos for research. To date, these attempts haven’t been fruitful.

Transgenic animals, SMGT, and CRISPR/Cas9

  • Researchers have increasingly shown interest in producing transgenic animals, i.e., animals carrying a gene that has been artificially inserted into their genome. This process is achieved through sperm mediated gene transfer (SMGT), where sperm cells are exposed to external DNA.
  • The successful creation of transgenic equine blastocysts has proven the potential of SMGT. However, it has its limitations; it can’t be used for specific applications like gene therapy because it doesn’t support the induction of targeted mutations.
  • The authors mention CRISPR/Cas9 technology as a vital tool for this purpose. CRISPR/Cas9 enables targeted modifications to the genome, making it a useful mechanism for various applications in genetic research, including gene therapy.

In their conclusion, the authors argue that parthenogenesis, SCNT, and interspecies SCNT are all forms of genetic manipulation, as they generate embryos that are genetically identical to their parent cell. They also illustrate the use of SMGT and CRISPR/Cas9 for direct modification of equine embryos.

Cite This Article

APA
Hisey EA, Ross PJ, Meyers S. (2021). Genetic Manipulation of the Equine Oocyte and Embryo. J Equine Vet Sci, 99, 103394. https://doi.org/10.1016/j.jevs.2021.103394

Publication

Journal of equine veterinary science
ISSN: 0737-0806
NlmUniqueID: 8216840
Country: United States
Language: English
Volume: 99
Pages: 103394

Researcher Affiliations

Hisey, Erin A
  • Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA.
Ross, Pablo J
  • Department of Animal Science, University of California, Davis, CA.
Meyers, Stuart
  • Department of Anatomy, Physiology, and Cell Biology, School of Veterinary Medicine, University of California, Davis, CA. Electronic address: smeyers@ucdavis.edu.

MeSH Terms

  • Animals
  • Blastocyst
  • Embryo, Mammalian
  • Horses
  • Male
  • Nuclear Transfer Techniques / veterinary
  • Oocytes
  • Parthenogenesis

Grant Funding

  • T32 GM136559 / NIGMS NIH HHS
  • T35 OD010956 / NIH HHS

Conflict of Interest Statement

Conflicts of Interest: None.

References

This article includes 76 references
  1. Shin MR, Cui XS, Jun JH, Jeong YJ, Kim NH. Identification of mouse blastocyst genes that are downregulated by double-stranded RNA-mediated knockdown of Oct-4 expression.. Mol Reprod Dev 2005;70(4):390–6.
    pubmed: 15685634
  2. Daigneault BW, Rajput S, Smith GW, Ross PJ. Embryonic POU5F1 is required for expanded bovine blastocyst formation.. Sci Rep 2018;8(1):7753.
    pmc: PMC5958112pubmed: 29773834
  3. Fogarty NME, McCarthy A, Snijders KE, Powell BE, Kubikova N, Blakeley P. Genome editing reveals a role for OCT4 in human embryogenesis.. Nature 2017;550(7674):67–73.
    pmc: PMC5815497pubmed: 28953884
  4. Yoon Y, Wang D, Tai PWL, Riley J, Gao G, Rivera-Pérez JA. Streamlined ex vivo and in vivo genome editing in mouse embryos using recombinant adeno-associated viruses.. Nat Commun 2018;9(1):412.
    pmc: PMC5788975pubmed: 29379011
  5. Jayakumar MK, Bansal A, Li BN, Zhang Y. Mesoporous silica-coated upconversion nanocrystals for near infrared light-triggered control of gene expression in zebrafish.. Nanomedicine (Lond) 2015;10(7):1051–61.
    pubmed: 25929564
  6. Gulías P, Guerra-Varela J, Gonzalez-Aparicio M, Ricobaraza A, Vales A, Gonzalez-Aseguinolaza G. Danio Rerio as model organism for adenoviral vector evaluation.. Genes (Basel) 2019;10(12):1053.
    doi: 10.3390/genes10121053pmc: PMC6947401pubmed: 31861246google scholar: lookup
  7. Zhang W, Aida T, Del Rosario RCH, Wilde JJ, Ding C, Zhang X. Multiplex precise base editing in cynomolgus monkeys.. Nat Commun 2020;11(1):2325.
    pmc: PMC7214463pubmed: 32393762
  8. Liang P, Xu Y, Zhang X, Ding C, Huang R, Zhang Z. CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes.. Protein Cell 2015;6(5):363–72.
    pmc: PMC4417674pubmed: 25894090
  9. Vincent MC, Trapnell BC, Baughman RP, Wert SE, Whitsett JA, Iwamoto HS. Adenovirus-mediated gene transfer to the respiratory tract of fetal sheep in utero.. Hum Gene Ther 1995;6(8):1019–28.
    pubmed: 7578414
  10. David AL, McIntosh J, Peebles DM, Cook T, Waddington S, Weisz B. Recombinant adeno-associated virus-mediated in utero gene transfer gives therapeutic transgene expression in the sheep.. Hum Gene Ther 2011;22(4):419–26.
    pubmed: 20919876
  11. Balakier H, Tarkowski AK. Diploid parthenogenetic mouse embryos produced by heat-shock and Cytochalasin B.. J Embryol Exp Morphol 1976;35(1):25–39.
    pubmed: 944760
  12. Hinrichs K, Schmidt AL, Selgrath JP. Activation of Horse Oocytes.. Biol Reprod Monograph Ser 1995;1:319–24.
  13. Brevini TA, Gandolfi F. Parthenotes as a source of embryonic stem cells.. Cell Prolif 2008;41(Suppl 1):20–30 Suppl 1.
    pmc: PMC6496533pubmed: 18181942
  14. Carneiro G, Lorenzo P, Pimentel C, Pegoraro L, Bertolini M, Ball B. Influence of insulin-like growth factor-I and its interaction with gonadotropins, estradiol, and fetal calf serum on in vitro maturation and parthenogenic development in equine oocytes.. Biol Reprod 2001;65(3):899–905.
    pubmed: 11514356
  15. Li X, Dai Y, Allen WR. Influence of insulin-like growth factor-I on cytoplasmic maturation of horse oocytes in vitro and organization of the first cell cycle following nuclear transfer and parthenogenesis.. Biol Reprod 2004;71(4):1391–6.
    pubmed: 15215202
  16. Choi YH, Velez IC, Macías-García B, Riera FL, Ballard CS, Hinrichs K. Effect of clinically-related factors on in vitro blastocyst development after equine ICSI.. Theriogenology 2016;85(7):1289–96.
    pubmed: 26777560
  17. Minamihashi A, Watson AJ, Watson PH, Church RB, Schultz GA. Bovine parthenogenetic blastocysts following in vitro maturation and oocyte activation with ethanol.. Theriogenology 1993;40(1):63–76.
    pubmed: 16727294
  18. Mitalipov SM, White KL, Farrar VR, Morrey J, Reed WA. Development of nuclear transfer and parthenogenetic rabbit embryos activated with inositol 1,4,5-trisphosphate.. Biol Reprod 1999;60(4):821–7.
    pubmed: 10084954
  19. Choi YH, Norris JD, Velez IC, Jacobson CC, Hartman DL, Hinrichs K. A viable foal obtained by equine somatic cell nuclear transfer using oocytes recovered from immature follicles of live mares.. Theriogenology 2013;79(5):791–6 e1.
    pubmed: 23312717
  20. Choi YH, Love CC, Varner DD, Thompson JA, Hinrichs K. Activation of cumulus-free equine oocytes: effect of maturation medium, calcium ionophore concentration and duration of cycloheximide exposure.. Reproduction 2001;122(1):177–83.
    pubmed: 11425342
  21. Bedford SJ, Kurokawa M, Hinrichs K, Fissore RA. Intracellular calcium oscillations and activation in horse oocytes injected with stallion sperm extracts or spermatozoa.. Reproduction 2003;126(4):489–99.
    pubmed: 14525531
  22. Choi YH, Love LB, Westhusin ME, Hinrichs K. Activation of equine nuclear transfer oocytes: methods and timing of treatment in relation to nuclear remodeling.. Biol Reprod 2004;70(1):46–53.
    pubmed: 12954733
  23. Choi YH, Love CC, Chung YG, Varner DD, Westhusin ME, Burghardt RC. Production of nuclear transfer horse embryos by Piezo-driven injection of somatic cell nuclei and activation with stallion sperm cytosolic extract.. Biol Reprod 2002;67(2):561–7.
    pubmed: 12135896
  24. Choi YH, Chung YG, Walker SC, Westhusin ME, Hinrichs K. In vitro development of equine nuclear transfer embryos: effects of oocyte maturation media and amino acid composition during embryo culture.. Zygote 2003;11(1):77–86.
    pubmed: 12625532
  25. Li X, Morris LH, Allen WR. Effects of different activation treatments on fertilization of horse oocytes by intracytoplasmic sperm injection.. J Reprod Fertil 2000;119(2):253–60.
    pubmed: 10864837
  26. Heras S, Smits K, Leemans B, Van Soom A. Asymmetric histone 3 methylation pattern between paternal and maternal pronuclei in equine zygotes.. Anal Biochem 2015;471:67–9.
    pubmed: 25461478
  27. Zhou H, Liu C, Wang W. Heterospecific nuclear-transferred embryos derived from equine fibroblast cells and enucleated bovine oocytes.. Reprod Domest Anim 2007;42(3):243–7.
    pubmed: 17506801
  28. Reggio B, Sansinena M, Cochran R, Guitreau A, Carter J, Denniston R. Nuclear transfer embryos in the horse.. In: Proceedings of the 5th International Symposium on Equine Embryo Transfer; 2001.
  29. Galli C, Colleoni S, Duchi R, Lagutina I, Lazzari G. Developmental competence of equine oocytes and embryos obtained by in vitro procedures ranging from in vitro maturation and ICSI to embryo culture, cryopreservation and somatic cell nuclear transfer.. Anim Reprod Sci 2007;98(1–2):39–55.
    pubmed: 17101246
  30. Choi YH, Hartman DL, Fissore RA, Bedford-Guaus SJ, Hinrichs K. Effect of sperm extract injection volume, injection of PLCzeta cRNA, and tissue cell line on efficiency of equine nuclear transfer.. Cloning Stem Cells 2009;11(2):301–8.
    pubmed: 19508114
  31. Hinrichs K, Shin T, Love C, Varner D, Westhusin M. Comparison of bovine and equine oocytes as host cytoplasts for equine nuclear transfer.. In: Proceedings of the 5th international symposium on equine embryo transfer Havemeyer Foundation Mono Ser; 2001.
  32. Woods GL, White KL, Vanderwall DK, Li GP, Aston KI, Bunch TD. A mule cloned from fetal cells by nuclear transfer.. Science 2003;301(5636):1063.
    pubmed: 12775846
  33. Galli C, Lagutina I, Crotti G, Colleoni S, Turini P, Ponderato N. Pregnancy: a cloned horse born to its dam twin.. Nature 2003;424(6949):635.
    pubmed: 12904778
  34. Hinrichs K. Assisted reproductive techniques in mares.. Reprod Domest Anim 2018;53(Suppl 2):4–13.
    pubmed: 30238661
  35. Lee RS, Peterson AJ, Donnison MJ, Ravelich S, Ledgard AM, Li N. Cloned cattle fetuses with the same nuclear genetics are more variable than contemporary half-siblings resulting from artificial insemination and exhibit fetal and placental growth deregulation even in the first trimester.. Biol Reprod 2004;70(1):1–11.
    pubmed: 13679311
  36. Constant F, Guillomot M, Heyman Y, Vignon X, Laigre P, Servely JL. Large offspring or large placenta syndrome? Morphometric analysis of late gestation bovine placentomes from somatic nuclear transfer pregnancies complicated by hydrallantois.. Biol Reprod 2006;75(1):122–30.
    pubmed: 16571872
  37. Johnson AK, Clark-Price SC, Choi YH, Hartman DL, Hinrichs K. Physical and clinicopathologic findings in foals derived by use of somatic cell nuclear transfer: 14 cases (2004–2008).. J Am Vet Med Assoc 2010;236(9):983–90.
    pubmed: 20433399
  38. Johnson AK, Hinrichs K. Neonatal care and management of foals derived by somatic cell nuclear transfer.. Methods Mol Biol 2015;1330:189–201.
    pubmed: 26621599
  39. Pozor MA, Sheppard B, Hinrichs K, Kelleman AA, Macpherson ML, Runcan E. Placental abnormalities in equine pregnancies generated by SCNT from one donor horse.. Theriogenology 2016;86(6):1573–82.
    pubmed: 27325574
  40. Lagutina I, Lazzari G, Duchi R, Turini P, Tessaro I, Brunetti D. Comparative aspects of somatic cell nuclear transfer with conventional and zona-free method in cattle, horse, pig and sheep.. Theriogenology 2007;67(1):90–8.
    pubmed: 17081599
  41. Hinrichs K, Choi YH, Varner DD, Hartman DL. Production of cloned horse foals using roscovitine-treated donor cells and activation with sperm extract and/or ionomycin.. Reproduction 2007;134(2):319–25.
    pubmed: 17660241
  42. Gambini A, Jarazo J, Olivera R, Salamone DF. Equine cloning: in vitro and in vivo development of aggregated embryos.. Biol Reprod 2012;87(1):15 1–9.
    pubmed: 22553223
  43. Gambini A, De Stéfano A, Jarazo J, Buemo C, Karlanian F, Salamone DF. Embryo aggregation does not improve the development of interspecies somatic cell nuclear transfer embryos in the horse.. Theriogenology 2016;86(4):1081–91.
    pubmed: 27157390
  44. Li X, Tremoleda JL, Allen WR. Effect of the number of passages of fetal and adult fibroblasts on nuclear remodelling and first embryonic division in reconstructed horse oocytes after nuclear transfer.. Reproduction 2003;125(4):535–42.
    pubmed: 12683924
  45. Vanderwall DK, Woods GL, Aston KI, Bunch TD, Li G, Meerdo LN. Cloned horse pregnancies produced using adult cumulus cells.. Reprod Fertil Dev 2004;16(7):675–9.
    pubmed: 15740690
  46. Olivera R, Moro LN, Jordan R, Luzzani C, Miriuka S, Radrizzani M. In vitro and in vivo development of horse cloned embryos generated with iPSCs, mesenchymal stromal cells and fetal or adult fibroblasts as nuclear donors.. PLoS One 2016;11(10):e0164049.
    pmc: PMC5061425pubmed: 27732616
  47. Olivera R, Moro LN, Jordan R, Pallarols N, Guglielminetti A, Luzzani C. Bone marrow mesenchymal stem cells as nuclear donors improve viability and health of cloned horses.. Stem Cells Cloning 2018;11:13–22.
    pmc: PMC5818860pubmed: 29497320
  48. Choi YH, Ritthaler J, Hinrichs K. Production of a mitochondrial-DNA identical cloned foal using oocytes recovered from immature follicles of selected mares.. Theriogenology 2014;82(3):411–17.
    pubmed: 24888683
  49. Gregg K, Polejaeva I. Risk of equine infectious anemia virus disease transmission through in vitro embryo production using somatic cell nuclear transfer.. Theriogenology 2009;72(3):289–99.
    pubmed: 19482352
  50. Asseged BD, Habtemariam T, Tameru B, Nganwa D. The risk of introduction of equine infectious anemia virus into USA via cloned horse embryos imported from Canada.. Theriogenology 2012;77(2):445–58.
    pmc: PMC3250577pubmed: 21958631
  51. Dominko T, Mitalipova M, Haley B, Beyhan Z, Memili E, McKusick B. Bovine oocyte cytoplasm supports development of embryos produced by nuclear transfer of somatic cell nuclei from various mammalian species.. Biol Reprod 1999;60(6):1496–502.
    pubmed: 10330111
  52. Tecirlioglu RT, Guo J, Trounson AO. Interspecies somatic cell nuclear transfer and preliminary data for horse-cow/mouse iSCNT.. Stem Cell Rev 2006;2(4):277–87.
    pubmed: 17848714
  53. Li GP, Seidel GE Jr, Squires EL. Interspecies cloning using fresh, store and dead equine and bovine somatic cells as donor nuclei and bovine cytoplasts.. Theriogenology 2002;57(1):432.
  54. Ball BA, Sabeur K, Allen WR. Liposome-mediated uptake of exogenous DNA by equine spermatozoa and applications in sperm-mediated gene transfer.. Equine Vet J 2008;40(1):76–82.
    pubmed: 18083664
  55. Smith K, Spadafora C. Sperm-mediated gene transfer: applications and implications.. Bioessays 2005;27(5):551–62.
    pubmed: 15832378
  56. Pereyra-Bonnet F, Fernández-Martín R, Olivera R, Jarazo J, Vichera G, Gibbons A. A unique method to produce transgenic embryos in ovine, porcine, feline, bovine and equine species.. Reprod Fertil Dev 2008;20(7):741–9.
    pubmed: 18842176
  57. Zaniboni A, Merlo B, Zannoni A, Bernardini C, Lavitrano M, Forni M. Expression of fluorescent reporter protein in equine embryos produced through intracytoplasmic sperm injection mediated gene transfer (ICSI-MGT).. Anim Reprod Sci 2013;137(1–2):53–61.
    pubmed: 23312469
  58. Webster NL, Forni M, Bacci ML, Giovannoni R, Razzini R, Fantinati P. Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer.. Mol Reprod Dev 2005;72(1):68–76.
    pubmed: 15906394
  59. Lavitrano M, Bacci ML, Forni M, Lazzereschi D, Di Stefano C, Fioretti D. Efficient production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation.. Proc Natl Acad Sci U S A 2002;99(22):14230–5.
    pmc: PMC137866pubmed: 12393815
  60. Lavitrano M, Camaioni A, Fazio VM, Dolci S, Farace MG, Spadafora C. Sperm cells as vectors for introducing foreign DNA into eggs: genetic transformation of mice.. Cell 1989;57(5):717–23.
    pubmed: 2720785
  61. Wang HJ, Lin AX, Zhang ZC, Chen YF. Expression of porcine growth hormone gene in transgenic rabbits as reported by green fluorescent protein.. Anim Biotechnol 2001;12(2):101–10.
    pubmed: 11808625
  62. Doudna JA, Charpentier E. Genome editing. The new frontier of genome engineering with CRISPR-Cas9.. Science 2014;346(6213):1258096.
    pubmed: 25430774
  63. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N. Multiplex genome engineering using CRISPR/Cas systems.. Science 2013;339(6121):819–23.
    pmc: PMC3795411pubmed: 23287718
  64. Jinek M, East A, Cheng A, Lin S, Ma E, Doudna J. RNA-programmed genome editing in human cells.. Elife 2013;2:e00471.
    pmc: PMC3557905pubmed: 23386978
  65. Mali P, Yang L, Esvelt KM, Aach J, Guell M, DiCarlo JE. RNA-guided human genome engineering via Cas9.. Science 2013;339(6121):823–6.
    pmc: PMC3712628pubmed: 23287722
  66. Chang N, Sun C, Gao L, Zhu D, Xu X, Zhu X. Genome editing with RNA-guided Cas9 nuclease in zebrafish embryos.. Cell Res 2013;23(4):465–72.
    pmc: PMC3616424pubmed: 23528705
  67. Blitz IL, Biesinger J, Xie X, Cho KW. Biallelic genome modification in F(0) Xenopus tropicalis embryos using the CRISPR/Cas system.. Genesis 2013;51(12):827–34.
    pmc: PMC4039559pubmed: 24123579
  68. Whitworth KM, Lee K, Benne JA, Beaton BP, Spate LD, Murphy SL. Use of the CRISPR/Cas9 system to produce genetically engineered pigs from in vitro-derived oocytes and embryos.. Biol Reprod 2014;91(3):78.
    pmc: PMC4435063pubmed: 25100712
  69. Crispo M, Mulet AP, Tesson L, Barrera N, Cuadro F, dos Santos-Neto PC. Efficient generation of myostatin knock-out sheep using CRISPR/Cas9 technology and microinjection into zygotes.. PLoS One 2015;10(8):e0136690.
    pmc: PMC4549068pubmed: 26305800
  70. Niu Y, Shen B, Cui Y, Chen Y, Wang J, Wang L. Generation of gene-modified cynomolgus monkey via Cas9/RNA-mediated gene targeting in one-cell embryos.. Cell 2014;156(4):836–43.
    pubmed: 24486104
  71. Pinzon-Arteaga C, Snyder MD, Lazzarotto CR, Moreno NF, Juras R, Raudsepp T. Efficient correction of a deleterious point mutation in primary horse fibroblasts with CRISPR-Cas9.. Sci Rep 2020;10(1):7411.
    pmc: PMC7198616pubmed: 32366884
  72. Hawkes JR. CRISPR/Cas9-mediated gene editing in HERDA equine.. Utah State University; 2019.
  73. Mançanares ACF, Cabezas J, Manríquez J, de Oliveira VC, Wong Alvaro YS, Rojas D. Edition of prostaglandin E2 receptors EP2 and EP4 by CRISPR/Cas9 technology in equine adipose mesenchymal stem cells.. Animals (Basel) 2020;10(6).
    pmc: PMC7341266pubmed: 32585798
  74. Vichera G, Viale D, Olivera R, Arnold V, Grundnig A, Baston J. Generation of myostatin knockout horse embryos using clustered regularly interspaced short palindromic repeats/CRISPR-associated gene 9 and somatic cell nuclear transfer.. Reprod Fertility Dev 2019;31(1):136.
  75. Moro LN, Viale DL, Bastón JI, Arnold V, Suvá M, Wiedenmann E. Generation of myostatin edited horse embryos using CRISPR/Cas9 technology and somatic cell nuclear transfer.. Sci Rep 2020;10(1):15587.
    pmc: PMC7518276pubmed: 32973188
  76. Navarro-Serna S, Vilarino M, Park I, Gadea J, Ross PJ. Livestock gene editing by one-step embryo manipulation.. J Equine Vet Sci 2020;89:103025.
    pubmed: 32563448

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