Abstract
High-throughput screening with a loss-of-function strategy is a logical and efficient way to identify novel genes involved in biological processes of interest. In zebrafish, morpholinos have been developed as a convenient tool to knock down gene expression. Here, we describe procedures for systematic screening using morpholinos in zebrafish to identify novel deubiquitylases involved in convergent extension during gastrulation. In this example, we examine candidates based on embryonic morphology and molecular signals of whole mount in situ hybridization assay.
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Acknowledgements
The authors are grateful to the Taiwan Zebrafish Core Facility in NHRI for their technical supports and fish maintenance. The deubiquitylase work is funded by the Ministry of Science and Technology, Taiwan, with a grant (MOST 103-2633-B-400-001) awarded to Y.J.J.
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Tseng, LC., Tang, CH., Jiang, YJ. (2016). Morphology and Gene Expression Screening with Morpholinos in Zebrafish Embryos. In: Azorsa, D., Arora, S. (eds) High-Throughput RNAi Screening. Methods in Molecular Biology, vol 1470. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6337-9_17
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DOI: https://doi.org/10.1007/978-1-4939-6337-9_17
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6335-5
Online ISBN: 978-1-4939-6337-9
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