To identify the protein partners of SARS-CoV-2 -1 PRF RNA, we performed RNA pull-down assays using streptavidin magnetic beads. To obtain more accurate and reliable data, tRSA sequence-tagged -1 PRF and -1 FSE RNA probes were both used to capture binding proteins in our screen. As a random sequence control for nonspecificity, the equivalent proteins extracted from H1299 cells were incubated with tRSA-labeled -1 PRF and -1 FSE RNA probes. A high-throughput technology, LC/MS-MS(MS), was applied to identify the candidate interacting proteins of SARS-CoV-2 -1 PRF RNA.