Genome-wide identification of protein binding sites on RNAs in mammalian cells [PBS-R]

We develop a novel method, called protein binding sites on RNA (PBS-R) to identify genome-wide protein binding sites on RNAs without using antibodies. Double click strategy is used for the PBS-R assay. Proteins and RNAs are UV-crosslinked in vivo first, then the proteins are crosslinked to the magnetic beads. The protein binding RNA elements are captured, reverse transcribed and sequenced. Overall design: Identification of RNA-protein binding patterns in Hela cells through high-throughput sequencing.