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. 2025 Jul 31;58(7):288–292. doi: 10.5483/BMBRep.2024-0102

Fig. 2.

Fig. 2

Cytosine base editing by five CBEs in protoplasts of A. thaliana and G. max. (A) The architecture of nCas9 (D10A)-fused cytidine deaminases utilized for C-to-T editing in protoplasts of A. thaliana and G. max is outlined. CBE, cytosine base editor; UGI, uracil DNA glycosylase inhibitor; nCas9, Cas9 nickase; Cde, cytidine deaminase; PAM, protospacer adjacent motif; gRNA, guide RNA; rAPOBEC1, rat APOBEC1; A3A, APOBEC3A; hAID, human AID; PmCDA1, Petromyzon marinus cytidine deaminase 1; YE1, rAPOBEC1/W90Y+R126E; NLS, nuclear localization sequence. (B) Five target sites (AtGL2, AtSKL1, AtVAR2, GmFT2a, and GmFT4) served to evaluate the editing efficiencies of various CBEs. Cytosines within the protospacer target sequence are highlighted in red. (C) Evaluations of five CBEs at three distinct target sites in A. thaliana protoplasts are documented. (D) The editing windows for five CBEs at three different target sites in A. thaliana protoplasts are delineated. (E) Cytosine base editing efficiencies of five CBEs at two independent target sites in G. max protoplasts are shown. (F) C-to-T editing windows of five CBEs within two target sites in G. max protoplasts are depicted. In (C, E), each dot signifies a biological replicate. In (C to F), the data are presented as means ± SEM. Distinct letters indicate a significant difference at P < 0.05 (one-way ANOVA with Tukey’s post hoc test).