Abstract
Here, we describe a fast and straightforward methodology to in vivo detect transcriptional activity in the early zebrafish germ line. We report how fluorescently labeled morpholinos, targeted to nascent early transcripts, can be used to track the onset of transcriptional events during early embryogenesis. This method could be applied to any tagged cell line in a developing early zebrafish embryo as long as the gene of interest is expressed at high enough level for morpholino detection and is expressed at the first and main wave of genome activation, for which the protocol has been verified. The protocol, in combination with genetic manipulation, allows studies of mechanisms driving zygotic genome activation (ZGA) in individual cells. The reported procedures apply to a broad range of purposes for zebrafish embryo manipulation in view of imaging nuclear molecules in specific cell types.
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Acknowledgments
This work was supported by the Horizon 2020 programme Zencode-ITN, Wellcome Trust Investigator award (106955/Z/15/Z) to F.M., Horizon 2020 FET project to F.M. We thank Roland Dosch at the University Medical Center in Göttingen for providing the Tg(Buc-GFP) line and the BMSU facility at the University of Birmingham for the maintenance.
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D’Orazio, F.M., Jasiulewicz, A., Hadzhiev, Y., Müller, F. (2021). Visualization of Transcriptional Activity in Early Zebrafish Primordial Germ Cells. In: Dosch, R. (eds) Germline Development in the Zebrafish. Methods in Molecular Biology, vol 2218. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0970-5_15
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DOI: https://doi.org/10.1007/978-1-0716-0970-5_15
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