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Functional Studies of Transcriptional Cofactors via Microinjection-Mediated Gene Editing in Xenopus

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Microinjection

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1874))

Abstract

The anuran Xenopus laevis has been studied for decades as a model for vertebrate cell and developmental biology. More recently, the highly related species Xenopus tropicalis has offered the opportunity to carry out genetic studies due to its diploid genome as compared to the pseudo-tetraploid Xenopus laevis. Amphibians undergo a biphasic development: embryogenesis to produce a free-living tadpoles and subsequent metamorphosis to transform the tadpole to a frog. This second phase mimics the so-called postembryonic development in mammals when many organs/tissues mature into their adult form in the presence of high levels of plasma thyroid hormone (T3). The total dependence of amphibian metamorphosis on T3 offers a unique opportunity to study postembryonic development in vertebrates, especially with the recent development gene editing technologies that function in amphibians. Here, we first review the basic molecular understanding of the regulation of Xenopus metamorphosis by T3 and T3 receptors (TRs), and then describe a detailed method to use CRISPR to knock out the TR-coactivator SRC3 (steroid receptor coactivator 3), a histone acetyltransferase, in order to study its involvement in gene regulation by T3 in vivo and Xenopus development.

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Acknowledgments

This work in the laboratory was supported by the Intramural Research Program of National Institute of Child Health and Human Development, National Institutes of Health. Y. Shibata was supported in part by a Japan Society for the Promotion of Science Research Fellowship for Japanese Biomedical and Behavioral Researchers at the National Institutes of Health.

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Authors and Affiliations

  1. Section on Molecular Morphogenesis, Eunice Kennedy Shriver National Institute of Child Health and Human Development (NICHD), National Institutes of Health (NIH), Bethesda, MD, USA

    Yuki Shibata, Lingyu Bao, Liezhen Fu & Yun-Bo Shi

  2. Department of Endocrinology, The First Affiliated Hospital of Xi’an Jiaotong University School of Medicine, Xi’an, Shaanxi, People’s Republic of China

    Lingyu Bao & Bingyin Shi

Authors
  1. Yuki Shibata
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  2. Lingyu Bao
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  3. Liezhen Fu
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  4. Bingyin Shi
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  5. Yun-Bo Shi
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Correspondence to Yun-Bo Shi .

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Editors and Affiliations

  1. National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Chengyu Liu

  2. National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD, USA

    Yubin Du

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Shibata, Y., Bao, L., Fu, L., Shi, B., Shi, YB. (2019). Functional Studies of Transcriptional Cofactors via Microinjection-Mediated Gene Editing in Xenopus. In: Liu, C., Du, Y. (eds) Microinjection. Methods in Molecular Biology, vol 1874. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8831-0_29

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  • DOI: https://doi.org/10.1007/978-1-4939-8831-0_29

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