PT - JOURNAL ARTICLE AU - Alexander, Michael J. AU - Budinger, G.R. Scott AU - Reyfman, Paul A. TI - Breathing fresh air into respiratory research with single-cell RNA sequencing AID - 10.1183/16000617.0060-2020 DP - 2020 Jun 30 TA - European Respiratory Review PG - 200060 VI - 29 IP - 156 4099 - https://publications.ersnet.org//content/29/156/200060.short 4100 - https://publications.ersnet.org//content/29/156/200060.full SO - EUROPEAN RESPIRATORY REVIEW2020 Jun 30; 29 AB - The complex cellular heterogeneity of the lung poses a unique challenge to researchers in the field. While the use of bulk RNA sequencing has become a ubiquitous technology in systems biology, the technique necessarily averages out individual contributions to the overall transcriptional landscape of a tissue. Single-cell RNA sequencing (scRNA-seq) provides a robust, unbiased survey of the transcriptome comparable to bulk RNA sequencing while preserving information on cellular heterogeneity. In just a few years since this technology was developed, scRNA-seq has already been adopted widely in respiratory research and has contributed to impressive advancements such as the discoveries of the pulmonary ionocyte and of a profibrotic macrophage population in pulmonary fibrosis. In this review, we discuss general technical considerations when considering the use of scRNA-seq and examine how leading investigators have applied the technology to gain novel insights into respiratory biology, from development to disease. In addition, we discuss the evolution of single-cell technologies with a focus on spatial and multi-omics approaches that promise to drive continued innovation in respiratory research.Single-cell RNA sequencing is being used more and more in respiratory research. This technology can be leveraged in different ways in studies of lung development, structure and function, and lung diseases from pulmonary fibrosis to asthma to lung cancer. https://bit.ly/36TdvJT