Haimon, Z;
Volaski, A;
Orthgiess, J;
Boura-Halfon, S;
Varol, D;
Shemer, A;
Yona, S;
... Jung, S; + view all
(2018)
Re-evaluating microglia expression profiles using RiboTag and cell isolation strategies.
Nature Immunology
, 19
(6)
pp. 636-644.
10.1038/s41590-018-0110-6.
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Abstract
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.
| Type: | Article |
|---|---|
| Title: | Re-evaluating microglia expression profiles using RiboTag and cell isolation strategies |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1038/s41590-018-0110-6 |
| Publisher version: | https://doi.org/10.1038/s41590-018-0110-6 |
| Language: | English |
| Additional information: | This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Immunology, MESSENGER-RNA, IN-VIVO, TISSUE MACROPHAGES, REVEALS, SEQ, TRANSCRIPTOME, FRACTALKINE, IDENTITIES, RETENTION, DYNAMICS |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Medical Sciences > Div of Medicine > Experimental and Translational Medicine |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10053195 |
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