Table 2.

Nucleotide sequence of primers designed for the insertion of miRNA target sequences into EV-A71 genome.

Primer Designation	Nucleotide sequences
F1 (Fwd 5′-NTR)	5′-TTAAAACAGCTGTGGGTTGTACCCAC-3′
R1 (Rev let-7a)	5′-TGAGGTAGTAGGTTGTATAGTTGTTTGATTGTATTGAGG-3′
F2 (Fwd let 7a)	5′-AACTATACAACCTACTACCTCAATGGGCTCACAGGTGTC-3′
F2: (Fwd let 7a-scr)	5′-AATTACACGACTTATTATTTGAATGGGCTCACAGGTGTC-3′
R2: (let 7a-scr):	5′-TATGGGACCCGCTGTATCCCTAAGGGTAGTAATGGCAG-3′
R2 (Rev miR124a)	5′-TAAGGCACGCGGTGAATGCCAAAGGGTAGTAATGGCAG-3′
F3 (Fwd miR-124a)	5′-TGGCATTCACCGCGTGCCTTAGGAAAGTTCGGCCAGC-3′
F3 (Fwd miR-124a-scr)	5′-TAGCGTTAACTGCATGGCGCAGGAAAGTTCGGCCAGC- 3′
R3 (Rev 3′-NTR)	5′-GCTATTCCGGTTATAACAAATTTACCCCCACTAG-3′

Nucleotides that are in bold refer to the two miRNA target sequences. Primers designated let-7a-scr or miR124a-scr were primers designed to incorporate scrambled nucleotides into target sequences to produce virus controls that had disrupted base pairing with endogenous miRNA whilst conserving the WT encoded amino acids.