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Chapitre D'ouvrage Année : 2019

Medium-Throughput RNA In Situ Hybridization of Serial Sections from Paraffin-Embedded Tissue Microarrays

Résumé

(m)RNA spatiotemporal pattern of distribution is of key importance to decipher gene function. In this post-genomic era, numerous transcriptomic studies are made publicly available, sometimes reaching a tissular resolution and even more rarely the cellular level. This “one tissue-numerous genes” information can be completed by the reverse “one gene-numerous tissues” picture through traditional RNA in situ hybridization (ISH). Here, we present a method including (1) principles of transcriptomic data mining to be performed prior and following ISH and (2) a detailed step-by-step medium-throughput ISH protocol performed on serial sections from tissue microarrays. In a recent work, we implemented this method for 39 selected genes studied by medium-throughput ISH complementing an existing tissue-specific transcriptomic dataset focused on the model plant Arabidopsis seed development kinetics (Francoz et al., Scientific Reports 6:24644, 2016). This full integration of ISH and transcriptomics demonstrated the complementarity of both techniques in terms of tissue/cell specificity, signal sensitivity, gene specificity, and spatiotemporal resolution.
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Dates et versions

hal-04397032 , version 1 (16-01-2024)

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Edith Francoz, Philippe Ranocha, Christophe Dunand, Vincent Burlat. Medium-Throughput RNA In Situ Hybridization of Serial Sections from Paraffin-Embedded Tissue Microarrays. Julia A. Chekanova; Hsiao-Lin V. Wang. Plant Long Non-Coding RNAs, 1933, Springer, pp.99-130, 2019, Methods in Molecular Biology, 978-1-4939-9044-3. ⟨10.1007/978-1-4939-9045-0_6⟩. ⟨hal-04397032⟩
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