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

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.

journal_name

Nat Immunol

journal_title

Nature immunology

authors

Haimon Z,Volaski A,Orthgiess J,Boura-Halfon S,Varol D,Shemer A,Yona S,Zuckerman B,David E,Chappell-Maor L,Bechmann I,Gericke M,Ulitsky I,Jung S

doi

10.1038/s41590-018-0110-6

subject

Has Abstract

pub_date

2018-06-01 00:00:00

pages

636-644

issue

6

eissn

1529-2908

issn

1529-2916

pii

10.1038/s41590-018-0110-6

journal_volume

19

pub_type

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