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Cattle Cloning by Somatic Cell Nuclear Transfer

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Somatic Cell Nuclear Transfer Technology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2647))

Abstract

Cloning by somatic cell Nuclear Transfer (SCNT) is a powerful technology capable of reprograming terminally differentiated cells to totipotency for generating whole animals or pluripotent stem cells for use in cell therapy, drug screening, and other biotechnological applications. However, the broad usage of SCNT remains limited due to its high cost and low efficiency in obtaining live and healthy offspring. In this chapter, we first briefly discuss the epigenetic constraints responsible for the low efficiency of SCNT and current attempts to overcome them. We then describe our bovine SCNT protocol for delivering live cloned calves and addressing basic questions about nuclear reprogramming. Other research groups can benefit from our basic protocol and build up on it to improve SCNT in the future. Strategies to correct or mitigate epigenetic errors (e.g., correcting imprinting loci, overexpression of demethylases, chromatin-modifying drugs) can integrate the protocol described here.

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Acknowledgments

The authors thank the staff and students at the Laboratory of Molecular Morphophysiology and Development. Juliano Rodrigues Sangalli is supported by São Paulo Research Foundation—FAPESP, grant number #2016/13416-9. Rafael Vilar Sampaio is supported by MITACS and L’Alliance Boviteq Inc. scholarship (FR39379). Tiago Henrique Camara De Bem is supported by São Paulo Research Foundation—FAPESP, grant number #2016/22790-1. Flávio Vieira Meirelles is supported by São Paulo Research Foundation—FAPESP, grant number #2013/08135-2, National Counsel of Technological and Scientific Development (CNPq) grant number 465539/2014-9 and CAPES. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Author notes

  1. Juliano Rodrigues Sangalli, Rafael Vilar Sampaio and Tiago Henrique Camara De Bem contributed equally with all other contributors.

Authors and Affiliations

  1. Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, SP, Brazil

    Juliano Rodrigues Sangalli, Rafael Vilar Sampaio, Tiago Henrique Camara De Bem & Flávio Vieira Meirelles

  2. Centre de Recherche en Reproduction et Fértilité, Faculty of Veterinary Medicine, University of Montreal, Saint-Hyacinthe, QC, Canada

    Rafael Vilar Sampaio & Lawrence Charles Smith

Authors
  1. Juliano Rodrigues Sangalli
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  2. Rafael Vilar Sampaio
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  3. Tiago Henrique Camara De Bem
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  4. Lawrence Charles Smith
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  5. Flávio Vieira Meirelles
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Corresponding author

Correspondence to Flávio Vieira Meirelles .

Editor information

Editors and Affiliations

  1. Chemical Biology Graduate Program, Federal University of São Paulo – UNIFESP, Campus Diadema, Diadema - SP, Brazil

    Marcelo Tigre Moura

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Sangalli, J.R., Sampaio, R.V., De Bem, T.H.C., Smith, L.C., Meirelles, F.V. (2023). Cattle Cloning by Somatic Cell Nuclear Transfer. In: Moura, M.T. (eds) Somatic Cell Nuclear Transfer Technology . Methods in Molecular Biology, vol 2647. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3064-8_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3064-8_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3063-1

  • Online ISBN: 978-1-0716-3064-8

  • eBook Packages: Springer Protocols

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