Generation of mutant mice by co-injection of two crRNAs with Cas9 mutants. (A) The
designs of the crRNAs in the mouse Bcr locus. The genomic structure
is indicated at the top. The black bars and boxes show introns and exons,
respectively. Blue and red lines show specific target sequences and common sequences
of crRNA, respectively. The letters with yellow boxes designate PAM sequences. The
blue curved lines indicate tracrRNA. Longitudinal blue bars show hydrogen bonds. (B)
The proportions of the genotypes of embryos obtained by crRNA1/tracrRNA/Cas9WT (the
same data set as shown in Fig. 1C),
crRNA1, and 2/tracrRNA/nCas9, and by crRNA1 and 2/tracrRNA/fCas9 injection,
respectively. The percentages calculated from the number of mutants relative to the
number of analyzed embryos. (C) The designs of the crRNAs for the mouse
Abl1 locus. The genomic structure is indicated at the top. The
black bars and boxes show introns and exons, respectively. Blue and red lines show
specific target sequences and common sequences of crRNAs, respectively. The letters
with yellow boxes designate PAM sequences. The blue curved lines indicate tracrRNA.
Longitudinal blue bars show hydrogen bonds. (D) Representative electropherograms of
embryos injected with crRNA3/crRNA4/tracrRNA and with nCas9 and fCas9, respectively.
Underlines indicate the target sequences of each crRNA.