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Re-evaluating microglia expression profiles using RiboTag and cell isolation strategies

Transcriptome profiling is widely used to infer functional states of specific cell types, as well as their responses to stimuli, to define contributions to physiology and pathophysiology. Focusing on microglia, the brain’s macrophages, we report here a side-by-side comparison of classical cell-sorting-based transcriptome sequencing and the ‘RiboTag’ method, which avoids cell retrieval from tissue context and yields translatome sequencing information. Conventional whole-cell microglial transcriptomes were found to be significantly tainted by artifacts introduced by tissue dissociation, cargo contamination and transcripts sequestered from ribosomes. Conversely, our data highlight the added value of RiboTag profiling for assessing the lineage accuracy of Cre recombinase expression in transgenic mice. Collectively, this study indicates method-based biases, reveals observer effects and establishes RiboTag-based translatome profiling as a valuable complement to standard sorting-based profiling strategies.

Authors:   Zhana Haimon; Alon Volaski; Johannes Orthgiess; Sigalit Boura-Halfon; Diana Varol; Anat Shemer; Simon Yona; Binyamin Zuckerman; Eyal David; Louise Chappell-Maor; Ingo Bechmann; Martin Gericke; Igor Ulitsky; Steffen Jung
Journal:   Nature Immunology
Year:   2018
Pages:   1
DOI:   10.1038/s41590-018-0110-6
Publication date:   18-May-2018
Facts, background information, dossiers
  • tissue
  • sequencing
  • ribosomes
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