Sequencing approaches that allowed for a better resolution of the transcriptome have been a major goal in the transcriptomics field since the development of RNA-seq techniques. While RNA-seq provides gene expression data from one entire sample in bulk, single-cell analysis allows for a better characterization of gene expression associated to specific cell types. Single-cell RNA-seq (scRNA-seq) is a reliable technique to unravel transcriptomic features of the tissues of interest dissociated at a single-cell level. The main feature of the single-cell technique is its ability to generate barcoded individual cells that allow for tracking the origin of thousands to millions of transcripts and reveal new cell types associated to diseases and different cell types and states. In this chapter, we discuss how scRNA-seq has become the gold standard to deepen the understanding of the gene expression with single-cell resolution.