Droplet-based forward genetic screening of astrocyte–microglia cross-talk

Michael A. Wheeler, Iain C. Clark, Hong Gyun Lee, Zhaorong Li, Mathias Linnerbauer, Joseph M. Rone, Manon Blain, Camilo Faust Akl, Gavin Piester, Federico Giovannoni, Marc Charabati, Joon Hyuk Lee, Yoon Chul Kye, Joshua Choi, Liliana M. Sanmarco, Lena Srun, Elizabeth N. Chung, Lucas E. Flausino, Brian M. Andersen, Veit RothhammerHiroshi Yano, Tomer Illouz, Stephanie E.J. Zandee, Carolin Daniel, David Artis, Marco Prinz, Adam R. Abate, Vijay K. Kuchroo, Jack P. Antel, Alexandre Prat, Francisco J. Quintana

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Cell–cell interactions in the central nervous system play important roles in neurologic diseases. However, little is known about the specific molecular pathways involved, and methods for their systematic identification are limited. Here, we developed a forward genetic screening platform that combines CRISPR-Cas9 perturbations, cell coculture in picoliter droplets, and microfluidic-based fluorescence-activated droplet sorting to identify mechanisms of cell–cell communication. We used SPEAC-seq (systematic perturbation of encapsulated associated cells followed by sequencing), in combination with in vivo genetic perturbations, to identify microglia-produced amphiregulin as a suppressor of disease-promoting astrocyte responses in multiple sclerosis preclinical models and clinical samples. Thus, SPEAC-seq enables the high-throughput systematic identification of cell–cell communication mechanisms.

Original languageEnglish
Pages (from-to)1023-1030
Number of pages8
JournalScience
Volume379
Issue number6636
DOIs
StatePublished - 10 Mar 2023
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2023 American Association for the Advancement of Science. All rights reserved.

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