Xtended Information Figure E5. Base editing efficiencies of further ABE5 variantsNature. Author manuscript; out there in PMC 2018 April 25.Gaudelli et al.Pagea, A to G base editing efficiencies in HEK293T cells at six human genomic target DNA web sites of two ABE3.1 variants with two pairs of mutations isolated from spectinomycin collection of the round five library. b, A to G base editing efficiencies in HEK293T cells at six human genomic target DNA websites of ABE5 variants with various linker lengths. See Extended Information Figure E1 for ABE genotypes and architectures. Values and error bars reflect the mean and s.d. of three independent biological replicates performed on diverse days.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptExtended Data Figure E6. Base editing efficiencies of ABE7 variants at 17 genomic sitesNature. Author manuscript; offered in PMC 2018 April 25.Gaudelli et al.PageA to G base editing efficiencies in HEK293T cells at 17 human genomic target DNA internet sites of ABE7.1-7.five (a), and ABE7.6-7.ten (b). See Extended Data Figure E1 for ABE genotypes and architectures. c, A to G base editing efficiencies in U2OS cells at six human genomic target DNA websites of ABE7.8-7.10. The decrease editing efficiencies observed in U2OS cells compared with HEK293T cells are consistent with transfection efficiency differences in between the two cell lines; we observed transfection efficiencies of 40?5 in U2OS cells under the circumstances utilized within this study, compared to 65?0 in HEK293T cells. Values and error bars reflect the imply and s.d. of three independent biological replicates performed on diverse days.Author Manuscript Author Manuscript Author Manuscript Author ManuscriptExtended Data Figure E7. Activity window of late-stage ABEsa, Relative A to G base editing efficiencies in HEK293T cells of late-stage ABEs at protospacer positions 1? in two human genomic DNA sites that together spot an A at each and every of these positions. Values are normalized towards the maximum observed efficiency at every of the two web sites for every single ABE = 1. b, Relative A to G base editing efficiencies in HEK293T cells of late-stage ABEs at protospacer positions 1?8 and 20 across all 19 human genomic DNA web sites tested. Values are normalized to the maximum observed efficiency at every single of theNature.1,8-Dihydroxynaphthalene site Author manuscript; offered in PMC 2018 April 25.6-Bromo-7-azaindole Purity Gaudelli et al.Page19 internet sites for each ABE = 1. For (a) and (b), values and error bars reflect the mean and s.d. of three independent biological replicates performed on unique days.PMID:26446225 Author Manuscript Author Manuscript Author Manuscript Author ManuscriptExtended Information Figure E8. Rounds of evolution and engineering improved ABE processivityThe calculated mean normalized linkage disequilibrium (LD) among nearby target As at 6 to 17 human genomic target DNA web-sites for one of the most active ABEs emerging from each round of evolution and engineering. Greater LD values indicate that an ABE is extra probably to edit an A if a nearby A within the exact same DNA strand (the same sequencing study) can also be edited. LD values are normalized from 0 to 1 so that you can be independent of editing efficiency. Values and error bars reflect the imply and s.d. of normalized LD values from three independent biological replicates performed on unique days.Nature. Author manuscript; offered in PMC 2018 April 25.Gaudelli et al.PageAuthor Manuscript Author Manuscript Author ManuscriptExtended Information Figure E9. Evaluation of cellular RNAs in ABE7.10-treated cells compar.