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Our structure-guided binding analyses, using recombinantly purified full-length human PCBP2 and viral 3C proteins, along with the crystal structures of the cloverleaf-3C complexes, offered further insights into how these viruses assemble RNP complexes for their genome replication within host cells. Moreover, we demonstrated that antisense oligonucleotides could block high-affinity PCBP2-binding sites, highlighting the potential to develop therapeutics targeting this replication platform for enteroviral infections. 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