Additional file 2: of Harnessing accurate non-homologous end joining for efficient precise deletion in CRISPR/Cas9-mediated genome editing

Figure S1. Comparison of accurate NHEJ in repair of Cas9-induced DSBs between human cells (HEK293 or U2OS) and mouse ES cells. Figure S2. The frequency of “Accurate”, “Deletion”, “Insertion” and “IndDel” in group I events from 71 endogenous gene loci. Figure S3. Nearly all of the + 1 insertions with no additional mutations at repair junctions were templated. Figure S4. Generation of + 1 templated insertions (TI) resulted from paired Cas9 cleavage with the W/W, W/C, C/W, or C/C orientation of paired PAMs. Figure S5. Nearly all of the + 1 templated insertions with no additional mutations could be predicted from paired Cas9 cleavage at the third and fourth base upstream of a PAM. Figure S6. Correlation between + 1 template insertions (TI) and out-of-frame mutations derived from either of the Common, Ideal, or Paired methods. Figure S7. Effect of accurate NHEJ with a frequency below or above 30% on the frequency of out-of-frame editing. Figure S8. Correlation between the out-of-frame editing frequency and the frequency of templated insertions (TI) at 50 genome sites. Figure S9. Comparison of the in-frame editing frequency induced by Common, Paired, and Ideal at 20 genome sites. Figure S10. Distributions of microhomology at repair junctions in 53BP1 wild-type, 53BP1ΔTudor, and 53BP1ΔOD mouse ES cells. Figure S11. Flowchart of the paired Cas9-gRNA protocol for genome editing that requires precise deletion of defined length. (PDF 4737 kb)