Fig. 2

Knockout efficiency can be further increased by combining duplex extension with disruption of the continuous sequence of Ts. a The effect of duplex extension when mutating the fourth T to an A in four sgRNAs. The raw data are shown in Figure S4 in Additional file 1. b The effect of mutation of Ts at the indicated positions to A, C, or G when also extending the duplex by 5 bp. The raw data are shown in Figure S5 in Additional file 1. The graphs represent biological repeats from one of three independent experiments with similar results, shown as mean ± standard deviation (n = 3). Significance was calculated using Student's t-test: *P < 0.05; **P < 0.01; ***P < 0.005; ****P < 0.001. M mutant