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

• DOI: 10.1126/science.abq4822
• Publication Date: 2023-03-29T13:22:11Z
• Authors: 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 Rothhammer; Hiroshi Yano; Tomer Illouz; Stephanie E. J. Z...; Carolin Daniel; David Artis; Marco Prinz; Adam R. Abate; Vijay K. Kuchroo; Jack P. Antel; Alexandre Prat; Francisco J. Quin...
• Keywords: Cell type; Genetic screen; Cell Sorting; Identification; High-Throughput Screening

Cell-cell interactions in the central nervous system play important roles neurologic diseases. However, little is known about 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 picoliter droplets, microfluidic-based fluorescence-activated droplet sorting to identify mechanisms of cell-cell communication. We used SPEAC-seq (systematic perturbation encapsulated associated cells followed by sequencing), combination with vivo microglia-produced amphiregulin as suppressor disease-promoting astrocyte responses multiple sclerosis preclinical models clinical samples. Thus, enables high-throughput communication mechanisms.