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. 2017 Oct;27(10):1289-1292.
doi: 10.1038/cr.2017.111. Epub 2017 Aug 29.

Enhanced base editing by co-expression of free uracil DNA glycosylase inhibitor

Lijie Wang  1   2   3 , Wei Xue  4 , Lei Yan  1   2   3   5 , Xiaosa Li  1   2   3 , Jia Wei  4 , Miaomiao Chen  1 , Jing Wu  1 , Bei Yang  5 , Li Yang  4 , Jia Chen  1
Affiliations

Enhanced base editing by co-expression of free uracil DNA glycosylase inhibitor

Lijie Wang et al. Cell Res. 2017 Oct.
No abstract available

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Figures

Figure 1
Figure 1
Enhanced base editing system. (A-I) Enhanced base editing by co-expressing BE3 and free UGI from separate vectors in 293FT cells. Schematic diagram illustrating the design of sgRNA, BE3 and UGI expression vectors (A). The indel frequency (B), the C-to-T editing frequency at the indicated position of the sgRNA target region (D), the ratio of desired C-to-T editing to unwanted indels (F) and the fractions of C-to-T, C-to-A and C-to-G substitutions (H) were individually determined at the specified genomic sites for the indicated conditions and plotted as follows: orange represents BE3, faint blue represents BE + low UGI, blue represents BE3 + medium UGI, dark blue represents BE3 + high UGI and black represents dCas9. The positions of edited Cs in the sgFANCF, sgSite2 and sgRNF2 target regions were indicated with the base distal from the PAM set as position 1. Statistical analyses highlighted the significant differences between BE3 (orange) and BE3 + high UGI (dark blue) in indel frequency (C), in C-to-T editing frequency at the indicated position within sgRNA target region (E), in the ratio of desired C-to-T editing to unwanted indels (G) and in the fraction of C-to-T substitution (I). (J-R) Enhanced base editing by eBE-S1 and eBE-S3 in 293FT cells. Schematic diagram illustrating the design of sgRNA, BE3, eBE-S1 and eBE-S3 expression vectors (J). The indel frequency (K), the C-to-T editing frequency (M), the ratio of desired C-to-T editing to unwanted indels (O) and the fractions of C-to-T, C-to-A and C-to-G substitutions (Q) were individually determined at the indicated genomic sites for BE3 (orange), eBE-S1(faint cyan) and eBE-S3 (cyan). The positions of edited Cs in the sgEMX1, sgFANCF, sgSite2, sgSite4 and sgRNF2 target regions were indicated with the base distal from the PAM set as position 1. Statistical analyses highlighted the significant differences between BE3 (orange) and eBE-S3 (cyan) in indel frequency (L), in the C-to-T editing frequency (N), in the ratio of desired C-to-T editing to unwanted indels (P) and in the fraction of C-to-T substitution (R). (B, D, F, K, M and O) Error bars (±), standard deviations of 3 replicates. (C, E, G, I, L, N, P and R) P values, one-tailed Student's t-test.
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