Fig. 1.

Representative results of Sanger sequencing of blastocysts formed after cytoplasmic microinjection with Cas9 protein and GalT gRNA, and the frequencies of indels decomposed from Sanger sequence data by TIDE analysis. (a) Analysis of wild-type blastocyst. (b) Analysis of a blastocyst carrying mosaic mutation. The Sanger sequence traces consist of a mixture of wild-type and 1 bp insertion sequence, which yields a composite sequence trace after the break site. TIDE analysis decomposed the Sanger sequence data, and indels and their frequencies are indicated. (c) Analysis of blastocyst carrying biallelic mutation. The Sanger sequence traces of a blastocyst carrying biallelic mutation consist of a composite sequence trace without wild-type sequence. Arrowheads indicate the Cas9 cleavage sites. Nucleotides in blue color indicate target sequences, and nucleotides in red color indicate protospacer adjacent motif (PAM) sequences.