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CRISPR/Cas9-Mediated Genetic Engineering to Generate a Disease Model Prairie Vole, Based on Species-Optimized Assisted Reproductive Technology

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Oxytocin

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2384))

Abstract

Social and prosocial behaviors, including communication, social bonding, and affiliation, parental behaviors, and empathy are key features of a highly social mammalian species. However, the neuronal mechanism in the brain underlying these behaviors remains unclear because of limited information on the social and prosocial behavioral levels in rodent models generally used in behavioral neuroscience studies.

The rodent species, prairie vole (Microtus ochrogaster), is one of the nontraditional animal models with several advantages in experimental science over other rodent models, such as mice or rats. Additionally, it demonstrates characteristics advantageous in the study of social and prosocial behaviors, such as monogamous pair bonding behavior, biparental care, and consoling behavior toward partners stressed by aversive foot shock stimulus. Recent studies of prairie voles have highlighted the importance of oxytocin (OXT) and oxytocin receptor (OXTR)-mediated mechanisms in the regulation of these behaviors.

Recently, we established assisted reproductive technologies for prairie voles, and successfully and efficiently generated an OXTR gene knockout (KO) prairie vole using Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR-associated protein 9 (Cas9 ), a powerful genome editing tool with artificially developed single-strand guide RNAs (sgRNA) and Cas9 endonucleases.

Herein, we describe the method for CRISPR /Cas9-mediated generation of OXTR KO prairie vole. This OXTR KO prairie vole can be a valuable tool to understand their unique social and prosocial behaviors and elucidate how the oxytocin system influences or modulates these behaviors in the brain.

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Acknowledgments

This research was supported by JSPS Grant-in-Aid for Scientific Research (A) [Grant Numbers 15H02442 (2015–2018)], JSPS Grant-in-Aid for challenging Exploratory Research [Grant Numbers 16K15698 (2016–2017)], JSPS Grant in Aid for JSPS Research Fellow [Grant Number 16J05070 (2016–2019)], and Strategic Research Program for Brain Sciences from Japan Agency for Medical Research and Development (AMED) [Grant Numbers 18dm0107076h0003 (2016–2020)].

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Authors and Affiliations

  1. Tohoku University Graduate School of Agricultural Science, Sendai, Miyagi, Japan

    Kengo Horie & Katsuhiko Nishimori

  2. Division of Behavioral Neuroscience and Psychiatric Disorders, Emory University, Atlanta, GA, USA

    Kengo Horie

  3. Department of Obesity and Internal Inflammation, Fukushima Medical University, Fukushima, Japan

    Katsuhiko Nishimori

Authors
  1. Kengo Horie
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  2. Katsuhiko Nishimori
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Correspondence to Katsuhiko Nishimori .

Editor information

Editors and Affiliations

  1. School of Chemistry, The University of Sydney, Camperdown, NSW, Australia

    Eryn L. Werry

  2. Research School of Chemistry, Australian National University, Canberra, ACT, Australia

    Tristan A. Reekie

  3. School of Chemistry, The University of Sydney, Camperdown, NSW, Australia

    Michael Kassiou

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Horie, K., Nishimori, K. (2022). CRISPR/Cas9-Mediated Genetic Engineering to Generate a Disease Model Prairie Vole, Based on Species-Optimized Assisted Reproductive Technology . In: Werry, E.L., Reekie, T.A., Kassiou, M. (eds) Oxytocin. Methods in Molecular Biology, vol 2384. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1759-5_9

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  • DOI: https://doi.org/10.1007/978-1-0716-1759-5_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1758-8

  • Online ISBN: 978-1-0716-1759-5

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