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

Wheeler MA, Clark IC, Lee HG, Li Z, Linnerbauer M, Rone JM, Blain M, Akl CF, Piester G, Giovannoni F, Charabati M, Lee JH, Kye YC, Choi J, Sanmarco LM, Srun L, Chung EN, Flausino LE, Andersen BM, Rothhammer V, Yano H, Illouz T, Zandee SE, Daniel C, Artis D, Prinz M, Abate AR, Kuchroo VK, Antel JP, Prat A, Quintana FJ (2023)

Publication Type: Journal article

Publication year: 2023

Journal

Science American Association for the Advancement of Science

Book Volume: 379

Pages Range: 1023-1030

Journal Issue: 6636

DOI: 10.1126/science.abq4822

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.

Involved external institutions

Veterans Affairs Healthcare System Boston and Harvard Medical School US United States (USA) (US) Montreal Neurological Institute and Hospital (MNI, The Neuro) CA Canada (CA) Weill Cornell Medicine US United States (USA) (US) Centre hospitalier de l'Université de Montréal (CHUM) CA Canada (CA) Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (HMGU) / Helmholtz Munich DE Germany (DE) Albert-Ludwigs-Universität Freiburg DE Germany (DE) University of California San Francisco (UCSF) US United States (USA) (US)

How to cite

APA:

Wheeler, M.A., Clark, I.C., Lee, H.G., Li, Z., Linnerbauer, M., Rone, J.M.,... Quintana, F.J. (2023). Droplet-based forward genetic screening of astrocyte-microglia cross-talk. Science, 379(6636), 1023-1030. https://dx.doi.org/10.1126/science.abq4822

MLA:

Wheeler, Michael A., et al. "Droplet-based forward genetic screening of astrocyte-microglia cross-talk." Science 379.6636 (2023): 1023-1030.

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