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In this work, small self-assembled messenger RNA nanospheres (mRNA-NSs) were successfully prepared by rolling circle transcription on a constructed apoptin plasmid. The self-assembled mRNA-NSs have a uniform diameter of approximately 65 nm, good dispersity in solution, and efficient therapeutic apoptin expression ability. In addition, the mRNA-NSs have a high loading capacity of 8.2% for the antitumor drug doxorubicin (Dox), which can effectively deliver the loaded Dox into 4 T1 cells. Cellular experiments show that Dox-loaded self-asse