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. 2021:2218:185-194.
doi: 10.1007/978-1-0716-0970-5_15.

Visualization of Transcriptional Activity in Early Zebrafish Primordial Germ Cells

Fabio M D'Orazio #  1   2   3 , Aleksandra Jasiulewicz #  1 , Yavor Hadzhiev  1 , Ferenc Müller  4
Affiliations

Visualization of Transcriptional Activity in Early Zebrafish Primordial Germ Cells

Fabio M D'Orazio et al. Methods Mol Biol. 2021.

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.

Keywords: Bucky ball; Germ cells; Germ plasm; Microinjections; Microscopy; Morpholinos; Transcription; ZGA; Zebrafish.

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References

    1. Reisser M, Palmer A, Popp AP, Jahn C, Weidinger G, Gebhardt JCM (2018) Single-molecule imaging correlates decreasing nuclear volume with increasing TF-chromatin associations during zebrafish development. Nat Commun 9(1):5218 - DOI
    1. Fukaya T, Lim B, Levine M (2016) Enhancer control of transcriptional bursting. Cell 166(2):358–368 - DOI
    1. Stapel LC, Broaddus C, Vastenhouw NL (2018) Detection and automated analysis of single transcripts at subcellular resolution in zebrafish embryos. Methods Mol Biol Clifton NJ 1649:143–162 - DOI
    1. Schulz KN, Harrison MM (2019) Mechanisms regulating zygotic genome activation. Nat Rev Genet 20(4):221–234 - DOI
    1. Chen J, Wu J, Hong Y (2016) The morpholino molecular beacon for specific RNA visualization in vivo. Chem Commun Camb Engl 52(15):3191–3194 - DOI

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