Discovery of plasma membrane associated RNAs through APEX-seq

In addition to nucleic acids, a variety of biomolecules have been found on the plasma membrane. Whereas, researchers have realized that RNA has the ability to bind to membrane vesicles in vitro and connect to the plasma membrane of the cell. Hence, the combination of high-throughput sequencing and in suit labelling methods provides approaches for large-scale identification of subcellular RNAs. Here, we applied the recently published method APEX-seq, and identified 75 plasma membrane-associated RNAs, of which lncRNA PMAR72 is located around the membrane. In addition, we observed that PMAR72 was concentrated into foci in specific membrane areas. Our findings will provide some new evidence to elaborate the relationship between RNA and the plasma membrane of mammalian cells. APEX-seq libraries for both labeled targets and unlabeled controls were generated for the following constructs:Lck(inside), Lyn(inside), CD28(outside), PDGFR(outside). Six to eight libraries were prepared for each construct (3 biological replicates for labeled targets, 3 biological replicates for unlabeled controls).