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Genetic Manipulation of the Avian Urogenital System Using In Ovo Electroporation

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Book cover Avian and Reptilian Developmental Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1650))

Abstract

One of the advantages of the avian embryo as an experimental model is its in ovo development and hence accessibility for genetic manipulation. Electroporation has been used extensively in the past to study gene function in chicken and quail embryos . Readily accessible tissues such as the neural tube, somites, and limb bud, in particular, have been targeted. However, more inaccessible tissues, such as the embryonic urogenital system , have proven more challenging to study. Here, we describe the use of in ovo electroporation of TOL2 vectors or RCASBP avian viral vectors for the rapid functional analysis of genes involved in avian sex determination and urogenital development . In the context of the developing urogenital system , these vectors have inherent advantages and disadvantages, which will be considered here. Either vector can both be used for mis-expressing a gene and for targeting endogenous gene knockdown via expression of short hairpin RNAs (shRNAs). Both of these vectors integrate into the genome and are hence spread throughout developing tissues. Going forward, electroporation could be combined with CRISPR/Cas9 technology for targeted genome editing in the avian urogenital system .

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Author information

Authors and Affiliations

  1. Department of Anatomy and Development Biology, Monash University, Clayton, VIC, 3800, USA

    Claire E. Hirst & Craig A. Smith

  2. Australian Regenerative Medicine Institute (ARMI), Monash University, Clayton, VIC, USA

    Olivier Serralbo

  3. Murdoch Children’s Research Institute, Royal Children’s Hospital, The University of Melbourne, Melbourne, VIC, Australia

    Katie L. Ayers & Kelly N. Roeszler

  4. Department of Paediatrics, Royal Children’s Hospital, The University of Melbourne, Melbourne, VIC, Australia

    Katie L. Ayers & Kelly N. Roeszler

Authors
  1. Claire E. Hirst
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  2. Olivier Serralbo
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  3. Katie L. Ayers
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  4. Kelly N. Roeszler
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  5. Craig A. Smith
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Corresponding author

Correspondence to Craig A. Smith .

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Editors and Affiliations

  1. IRCMS, Kumamoto University, Kumamoto, Kumamoto, Japan

    Guojun Sheng

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© 2017 Springer Science+Business Media LLC

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Hirst, C.E., Serralbo, O., Ayers, K.L., Roeszler, K.N., Smith, C.A. (2017). Genetic Manipulation of the Avian Urogenital System Using In Ovo Electroporation. In: Sheng, G. (eds) Avian and Reptilian Developmental Biology. Methods in Molecular Biology, vol 1650. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7216-6_11

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  • DOI: https://doi.org/10.1007/978-1-4939-7216-6_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7215-9

  • Online ISBN: 978-1-4939-7216-6

  • eBook Packages: Springer Protocols

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