Project description:Several classes of small RNAs have been described, however to molecularly profile them require large numbers of cells. Here, we developed a method for single-cell small-RNA sequencing that we applied to naïve and primed human embryonic stem cells and to cancer cells. Single-cell profiling of microRNAs and fragments of tRNAs and snoRNAs revealed that, in particular, microRNAs have unrecognized potential to separate cell types and states.
Project description:miRNA and other forms of small RNA are now known to regulate many biological processes, and miRNA profiling is already used biomedically for cancer diagnosis, staging, progression, prognosis, and to evaluate responsivity to treatment. However, there is no robust analytical method capable of dissecting the individual miRNA expression profiles from the highly heterogeneous cells comprising tumor samples. Here, we developed parallel single cell small RNA sequencing (PSCSR-seq), which enables highly sensitive and high-throughput miRNA expression profiling of individual cells. We applied PSCSR-seq to examine the small RNA profiles from large numbers of cultured cancer cells, human PBMCs, and from a tumor in a melanoma mouse model. We anticipate that PSCSR-seq can be broadly applied for small RNA analysis in cancer research and more generally in life science.