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Gene Editing in Channel Catfish via Double Electroporation of Zinc-Finger Nucleases

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Zinc Finger Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1867))

Abstract

The traditional approach for gene editing with zinc-finger nucleases (ZFNs) in fish has been microinjection of mRNA. Here, we develop and describe an alternative protocol in which ZFN plasmids are electroporated to channel catfish, Ictalurus punctatus, sperm, and embryos. Briefly, plasmids were propagated to supply a sufficient quantity for electroporation. Sperm cells were prepared in saline solution, electroporated with plasmids, and then used for fertilization. Embryos were incubated with the plasmids before performing electroporation just prior to first cell division. Plasmids were then transcribed and translated by embryonic cells to produce ZFNs for gene editing, resulting in mutated fry.

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Author information

Authors and Affiliations

  1. School of Fisheries Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, USA

    Rex A. Dunham, Ahmed Elaswad & Zhenkui Qin

  2. Department of Animal Wealth Development, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt

    Ahmed Elaswad

  3. Key Laboratory of Marine Genetics and Breeding (Ocean University of China), Ministry of Education, Qingdao, China

    Zhenkui Qin

Authors
  1. Rex A. Dunham
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  2. Ahmed Elaswad
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  3. Zhenkui Qin
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Corresponding author

Correspondence to Rex A. Dunham .

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Editors and Affiliations

  1. Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University, Shanghai, China

    Jia Liu

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Dunham, R.A., Elaswad, A., Qin, Z. (2018). Gene Editing in Channel Catfish via Double Electroporation of Zinc-Finger Nucleases. In: Liu, J. (eds) Zinc Finger Proteins. Methods in Molecular Biology, vol 1867. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8799-3_15

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  • DOI: https://doi.org/10.1007/978-1-4939-8799-3_15

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8798-6

  • Online ISBN: 978-1-4939-8799-3

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