. 2014 Sep 12;56(7):499–510. doi: 10.1111/dgd.12149

Table 5.

Mutation frequency by sgRNAs that contain single base mismatches at the recognition site

Target gene	Name of sgRNA	Method of introduction	Sequence of the recognition site in RNA^†	Mutation frequency (in %)	n^‡
Ci-Hox5	wt Hox5-sg1^§	Microinjection of 3.0 pg of sgRNA and 15 pg of Cas9 mRNA	GGCGACGACGGGUUAGGUAA	75.9^¶	31
			Mismatch1		GGCGcCGACGGGUUAGGUAA	3.3	30
			Mismatch2		GGCGACGcCGGGUUAGGUAA	0	31
			Mismatch3		GGCGACGACGuGUUAGGUAA	0	28
			Mismatch4		GGCGACGACGGGUgAGGUAA	6.7	29
			Mismatch5		GGCGACGACGGGUUAGuUAA	0	30
			Mismatch6		GGCGACGACGGGUUAGGUAc	0	31
Ci-Hox3	wt Hox3-sg3^§	Electroporation of 20 μg of sgRNA and 40 μg of Cas9 expression vectors	GGCAGCCAUAAGAGUCAACA	36.3^¶	22
			Mismatch1		GGCAGCCcUAAGAGUCAACA	0	23
			Mismatch2		GGCAGCCAUAcGAGUCAACA	13.6	22
			Mismatch3		GGCAGCCAUAAGAuUCAACA	0	24
			Mismatch4		GGCAGCCAUAAGAGUCcACA	0	24
			Mismatch5		GGCAGCCAUAAGAGUCAACc	0	24

†Mismatched nucleotides are shown in lower case. ‡Number of sequenced polymerase chain reaction (PCR) clones. §wild type (wt) indicating perfect matching sgRNA. ¶The scores are the same as those shown in Tables2 and 3.