Small RNA sequencing of cell RNA and extracellular vesicle (EV) RNA from MDA-MB-231-derived isogenic cells

To identify the cell miRNAs and EV miRNAs that are differentially expressed or secreted by the MDA-MB-231 metastatic breast cancer cells with genetic modifications, we profiled the small RNAs (between 17 and 52 nt) in cells and EVs by Illumina sequencing. miRNAs that are significantly altered by Rab27a knockdown (KD) or miR-122 knockout (KO) are identified by comparing to the control cells (CTRL). Overall design: EVs were collected by ultracentrifugation of the conditioned medium of MDA-MB-231-derived cells. Cell RNA was extracted using TRIZOL, whereas EV RNA was extracted using TRIZOL LS (Thermo Fisher Scientific), and subjected to library construction and Illumina sequencing.