| A preparation method of self-assembling nanoparticle includes the following steps. A first donor plasmid, a second donor plasmid and a third donor plasmid are provided. A transposition step of plasmids is performed. A preparation step of recombinant viruses is performed so as to obtain a first recombinant baculovirus, a second recombinant baculovirus and a third recombinant baculovirus. A transduction step is performed, wherein a producer cell is simultaneously infected by the first recombinant baculovirus, the second recombinant baculovirus and the third recombinant baculovirus, and then a nucleocapsid protein, an envelope protein, an engineered envelope protein and a target RNA are expressed from the producer cell, and the nucleocapsid protein, the envelope protein, the engineered envelope protein and the target RNA self-assemble to form a self-assembling nanoparticle. Therefore, the production efficiency and the transfection efficiency of the self-assembling nanoparticle can be effectively enhanced, and the self-assembling nanoparticle can have excellent cell-targeting property. |