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We chemically synthesized RNA oligonucleotides containing BBV and gene of interest (GOI) with characteristic 5\u2019-OH and 3\u2019-P termini, which could undergo circularization by endogenous RtcB RNA ligases and efficiently encode proteins through RCT in mammalian cells. We designated these RNA oligonucleotides as Protein-Encoding RNA Oligonucleotides (PEOs). Notably, compared with IVT-produced RNAs, PEOs contained undetectable levels of proinflammatory double-stranded RNAs and exhibited minimal immunogenicity. We further demonstrated that PEO-OVA (encoding OVA antigens) significantly inhibited tumor growth comparable to mRNA vaccine. In an orthotopic glioma model, PEO vaccine also exhibited therapeutic benefits with checkpoint blockade therapy. 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