Figure 4 | Scientific Reports

Figure 4

From: Effective CRISPR/Cas9-mediated correction of a Fanconi anemia defect by error-prone end joining or templated repair

Figure 4

CRISPR/Cas9 templated gene editing analysis in mouse ES cells. (A) Overall gene editing efficiencies obtained in mouse embryonic stem cells depicted as general indel frequencies determined by TIDER (n = 3, one sided Student’s T-test, *p ≤ 0.02). (B) TIDE analysis of indel formation at predicted off-target loci after wtCas9 and sgRNA + 1 exposure irrespective of the ssODN orientation (n = 3, one sided Student’s T-test, *p ≤ 0.009). (C) Restriction fragment length polymorphism (RFLP) analysis by KpnI digest of the Fancf locus provides evidence for template-based repair after Cas9 transfection (n = 6, one sided Student’s T-test, *p ≤ 0.02). (D) Mouse ES cells show enhanced clonal survival in the presence of 12.5 nM MMC after double-strand break formation by Cas9 and sgRNA + 1 independent of the orientation of the HDR template. A significant difference in relative mESC clone survival was also observed with the antisense (AS) ssODN following Cas9 nickase activity (n = 6, one sided Student’s T-test, *p ≤ 0.025), “M” = mock sgRNA.

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