Chinmayee Goda, Shaopeng Gu, Anjun Ma, Mario Acunzo, Giulia Romano, Adrienne M Dorrance, Ramiro Garzon, Qin Ma, Giovanni Nigita
MicroRNAs (miRNAs) are small non-coding RNAs that play essential roles in gene regulation, cellular function, and disease pathogenesis. Advances in single-cell RNA sequencing technologies have enabled the profiling of miRNAs at single-cell resolution, providing unprecedented insight into cell-specific regulatory networks and heterogeneity. This chapter presents an overview of miRNA biology, technical approaches for single-cell miRNA sequencing, and recent bioinformatics tools developed for data analysis. We discuss challenges in library preparation, such as adapter biases and low RNA input, and highlight integrative strategies for co-profiling miRNAs with other omics layers. Finally, we outline in the conclusion the potential of single-cell miRNA profiling to contribute to precision medicine and therapeutic development, including its possible use in biomarker discovery, monitoring tumor heterogeneity, and informing personalized treatment strategies. As the field progresses, continued innovation will be critical to overcoming existing barriers and fully harnessing the power of single-cell miRNA analyses.