A full-length cDNA clone of X. laevis Dse.S in pCS105 (ID: XL487g09ex) that contained 251 nucleotides of the 5′ untranslated region (UTR), the open reading frame (ORF) and 680 nucleotides of the 3′UTR was obtained from the Osada/Taira NBRP Xenopus ANE library (Osada et al., 2003) and completely sequenced (GenBank accession number KU877109). To obtain pCS2-Dse*, the ORF of Dse.S was point-mutated by performing PCR and subcloned into pCS2, which resulted in seven nucleotide mismatches in the Dse-MO target sequence in positions −2, −1, +6, +9, +12, +15 and +18 relative to the translation start site. A full-length cDNA clone of X. laevis Dsel.L with 93 nucleotides of the 5′UTR, the ORF and 22 nucleotides of the 3′UTR (GenBank accession number KU877110) was generated by RT-PCR and subcloned into the pCR-XL-TOPO vector (Thermo Fisher Scientific), using total RNA from the head of stage 28 embryos and primers based on a corresponding Xenbase sequence entry.
