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

First Author  Haimon Z Year  2018
Journal  Nat Immunol Volume  19
Issue  6 Pages  636-644
PubMed ID  29777220 Mgi Jnum  J:282539
Mgi Id  MGI:6381230 Doi  10.1038/s41590-018-0110-6
Citation  Haimon Z, et al. (2018) Re-evaluating microglia expression profiles using RiboTag and cell isolation strategies. Nat Immunol 19(6):636-644
abstractText  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.
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