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Artikel ; Online: Establishing Riboglow-FLIM to visualize noncoding RNAs inside live zebrafish embryos.

Sarfraz, Nadia / Lee, Harrison J / Rice, Morgan K / Moscoso, Emilia / Shafik, Luke K / Glasgow, Eric / Ranjit, Suman / Lambeck, Ben J / Braselmann, Esther

Biophysical reports

2023  Band 3, Heft 4, Seite(n) 100132

Abstract: The central role of RNAs in health and disease calls for robust tools to visualize RNAs in living systems through fluorescence microscopy. Live zebrafish embryos are a popular system to investigate multicellular complexity as disease models. However, RNA ...

Abstract The central role of RNAs in health and disease calls for robust tools to visualize RNAs in living systems through fluorescence microscopy. Live zebrafish embryos are a popular system to investigate multicellular complexity as disease models. However, RNA visualization approaches in whole organisms are notably underdeveloped. Here, we establish our RNA tagging and imaging platform Riboglow-FLIM for complex cellular imaging applications by systematically evaluating FLIM capabilities. We use adherent mammalian cells as models for RNA visualization. Additional complexity of analyzing RNAs in whole mammalian animals is achieved by injecting these cells into a zebrafish embryo system for cell-by-cell analysis in this model of multicellularity. We first evaluate all variable elements of Riboglow-FLIM quantitatively before assessing optimal use in whole animals. In this way, we demonstrate that a model noncoding RNA can be detected robustly and quantitatively inside live zebrafish embryos using a far-red Cy5-based variant of the Riboglow platform. We can clearly resolve cell-to-cell heterogeneity of different RNA populations by this methodology, promising applicability in diverse fields.
Sprache Englisch
Erscheinungsdatum 2023-09-26
Erscheinungsland United States
Dokumenttyp Journal Article
ISSN 2667-0747
ISSN (online) 2667-0747
DOI 10.1016/j.bpr.2023.100132
Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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