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Article Dans Une Revue Cell Reports Methods Année : 2021

Interrogating RNA and protein spatial subcellular distribution in smFISH data with DypFISH

Nicolas Beaume
Edgar Gomes

Résumé

Advances in single-cell RNA sequencing have allowed for the identification of cellular subtypes on the basis of quantification of the number of transcripts in each cell. However, cells might also differ in the spatial distribution of molecules, including RNAs. Here, we present DypFISH, an approach to quantitatively investigate the subcellular localization of RNA and protein. We introduce a range of analytical techniques to interrogate single-molecule RNA fluorescence in situ hybridization (smFISH) data in combination with protein immunolabeling. DypFISH is suited to study patterns of clustering of molecules, the association of mRNA-protein subcellular localization with microtubule organizing center orientation, and interdependence of mRNA-protein spatial distributions. We showcase how our analytical tools can achieve biological insights by utilizing cell micropatterning to constrain cellular architecture, which leads to reduction in subcellular mRNA distribution variation, allowing for the characterization of their localization patterns. Furthermore, we show that our method can be applied to physiological systems such as skeletal muscle fibers.
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Origine : Publication financée par une institution

Dates et versions

hal-03349483 , version 1 (20-09-2021)

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Paternité - Pas d'utilisation commerciale - Pas de modification

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Anca Savulescu, Robyn Brackin, Emmanuel Bouilhol, Benjamin Dartigues, Jonathan Warrell, et al.. Interrogating RNA and protein spatial subcellular distribution in smFISH data with DypFISH. Cell Reports Methods, 2021, 1 (5), pp.100068. ⟨10.1016/j.crmeth.2021.100068⟩. ⟨hal-03349483⟩
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