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    • 4. 发明申请
    • RNAYLATION
    • WO2023006264A1
    • 2023-02-02
    • PCT/EP2022/060525
    • 2022-04-21
    • MAX-PLANCK-GESELLSCHAFT ZUR FÖRDERUNG DER WISSENSCHAFTEN E.V.UNIVERSITÄT HEIDELBERG
    • HÖFER, KatharinaJÄSCHKE, Andres
    • C12N9/10C12Q1/48A61P35/00C07K2319/00C07K2319/30C07K2319/70C12N9/1077C12Y204/0203
    • The present invention relates to a method for attaching a 5'-nicotinamidnucleobasedinucleotide (NND)- capped nucleic acid sequence to a fusion protein or to a complex, comprising (a) contacting (i) a heterologous fusion protein which comprises a poly(peptide) of interest being fused to a tag, or (ii) a complex wherein a protein is under physiological conditions complexed with a tag with the 5'-NAD- capped nucleic acid sequence and an ADP-ribosyltransferase (ART) under conditions wherein the 5'- NND-capped nucleic acid sequence is covalently attached to the tag, wherein the tag comprises a recognition motif of the ART and preferably comprises or consists of (i) SEQ ID NO: 1 or a sequence being at least 80% identical thereto provided that the underlined Arg in the amino acid motif DVRPVRD (SEQ ID NO: 7) is conserved and preferably SEQ ID NO: 7 is conserved; (ii) SEQ ID NO: 2 ora sequence being at least 80% identical thereto provided that the underlined Arg in the amino acid motif DVRPVRD (SEQ ID NO: 7) is conserved and preferably SEQ ID NO: 7 is conserved; (iii) SEQ ID NO: 3 or a sequence being at least 80% identical thereto provided that the underlined Arg in the amino acid motif DVRPVRD (SEQ ID NO: 7) is conserved and preferably SEQ ID NO: 7 is conserved; (iv) SEQ ID NO: 4 or a sequence being at least 80% identical thereto provided that the underlined Arg in the amino acid motif LADGVEGYLRASEASRDRVE (SEQ ID NO: 8) is conserved and preferably SEQ ID NO: 8 is conserved; (v) SEQ ID NO: 5 or a sequence being at least 80% identical thereto provided that the underlined Arg in the amino acid motif LADGVEGYLRASEASRDRVE (SEQ ID NO: 8) is conserved and preferably SEQ ID NO: 8 is conserved; or (vi) SEQ ID NO: 6 or a sequence being at least 80% identical thereto provided that the underlined Arg in the amino acid motif LADGVEGYLRASEASRDRVE (SEQ ID NO: 8) is conserved and preferably SEQ ID NO: 8 is conserved.