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. Author manuscript; available in PMC: 2020 Apr 1.
Published in final edited form as: Int J Parasitol. 2019 Feb 14;49(5):397–406. doi: 10.1016/j.ijpara.2018.12.004

Fig. 4.

Fig. 4.

An introduced F167Y mutation imparts thiabendazole resistance on wild type Caenorhabditis elegans. CRISPR/Cas9 was used to introduce the T to A mutation in the corresponding codon of the C. elegans ben-1 gene. (A) Alignment of predicted protein sequences in the region surrounding amino acid 167 in Ancylostoma caninum TBB-iso-1 and Caenorhabditis elegans BEN-1 proteins. Amino acid 167 is in bold, and asterisks indicate identical amino acids. (B) Diagram of the CRISPR/Cas9 strategy. The black arrow indicates the protospacer adjacent motif (PAM) site, and the white arrow indicates the antisense CRISPR RNA. The donor template (ssODN) is shown, with the changes in codon 167 and the PAM site indicated in bold italics. (C) Sequences of the CRISPR/Cas9 targeted region of eight independent lines of C. elegans. The first sequence represents the wild type DNA sequence, and the second sequence is the expected mutation in codon 167 and the targeted PAM site. Codon 167 and the PAM site are overlined. Bases that differ from the wild type are shown in bold, and asterisks indicate bases identical to the wild type ben-1 sequence.