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Using Morpholinos to Probe Gene Networks in Sea Urchin

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Morpholino Oligomers

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1565))

Abstract

The control processes that underlie the progression of development can be summarized in maps of gene regulatory networks (GRNs). A critical step in their assembly is the systematic perturbation of network candidates. In sea urchins the most important method for interfering with expression in a gene-specific way is application of morpholino antisense oligonucleotides (MOs). MOs act by binding to their sequence complement in transcripts resulting in a block in translation or a change in splicing and thus result in a loss of function. Despite the tremendous success of this technology, recent comparisons to mutants generated by genome editing have led to renewed criticism and challenged its reliability. As with all methods based on sequence recognition, MOs are prone to off-target binding that may result in phenotypes that are erroneously ascribed to the loss of the intended target. However, the slow progression of development in sea urchins has enabled extremely detailed studies of gene activity in the embryo. This wealth of knowledge paired with the simplicity of the sea urchin embryo enables careful analysis of MO phenotypes through a variety of methods that do not rely on terminal phenotypes. This article summarizes the use of MOs in probing GRNs and the steps that should be taken to assure their specificity.

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Acknowledgements

I am grateful to the many members of the sea urchin community who shared their experiences. Many thanks to Drs. Jonathan Rast, Eric Erkenbrack, Lauren Shipp, and Stephanie Woo for detailed comments on the manuscript. In memory of Eric Davidson.

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Authors and Affiliations

  1. School of Natural Sciences, University of California Merced, 5200 N. Lake Road, Merced, CA, 95340, USA

    Stefan C. Materna

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  1. Stefan C. Materna
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Correspondence to Stefan C. Materna .

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Editors and Affiliations

  1. Department of Biomedical Sciences College of Veterinary Medicine, Oregon State University, Corvallis, Oregon, USA

    Hong M. Moulton

  2. Gene Tools, LLC, Philomath, Oregon, USA

    Jon D. Moulton

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Materna, S.C. (2017). Using Morpholinos to Probe Gene Networks in Sea Urchin. In: Moulton, H., Moulton, J. (eds) Morpholino Oligomers. Methods in Molecular Biology, vol 1565. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6817-6_8

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  • DOI: https://doi.org/10.1007/978-1-4939-6817-6_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6815-2

  • Online ISBN: 978-1-4939-6817-6

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