Fig 2.

Mutations and deletion resulting from siRNA selection in vitro. (A) siRNAs used in this study: siNPPRV1 (lines 2 to 6), siNPPRV1mut4 (line 7), siNPPRV6 (lines 8 to 11), siNPPRV7 (lines 12 to 16), siNPPRV1 and 7 (line 17), siNPPRV1 and -6 (line 18), siNPPRV6 and -7 (line 19), and siNPPRV1, -6, and -7 (line 20). Line 1 shows the original nucleotide sequence of PPRV vaccine strain Nigeria 75/1 (underlined letters show the exact siRNA targets). No modification was found after 20 consecutive transfections in vitro when three siRNAs were combined (line 20). The column “passage” indicates the number of consecutive transfections when a modification was first detected. (B) Amino acid mutations in the siRNA-targeted regions are shown below the original sequences. Subscripts in the original sequences indicate the number of possible codons for the corresponding amino acid. The gray areas indicate the amino acids where all modifications occurred (synonymous or nonsynonymous mutations and deletion).