Fig. 3

Introduction of directed mutations into the amdS gene in M. thermophila by sgRNA-guided Cas9. a Schematic illustration of amdS mutagenesis by the CRISPR/Cas9 system. The M1 strain carrying the amdS gene was co-transformed with U6p-amdS-sgRNA and Cas9 cassettes, and then positive transformants were selected on medium plates containing 2 mg/mL FAA. b Sequence alignment of the amdS target locus from FAA-resistant transformants. The wild-type target sequence of amdS is framed in blue. Red letters depict the protospacer adjacent motif (PAM)