Fig. 4 | Scientific Reports

Fig. 4

From: Establishment and rescue of fibroblast cell lines carrying a nonsense mutation of RB1 by CRISPR-based base editing

Fig. 4

Rescue of the disrupted RB1 gene by adenine base editor (ABE)-mediated base editing in R552X cell lines. (A) Schematic of the ABE, ABEmax. Adenine deaminases, wild-type and evolved (*) TadA dimer, fused to nCas9 (D10A) and it converts an exposed adenosine into a guanosine. In the case of ABE8e, only an additionally engineered TadA* monomer is fused to nCas9. (B) Schematic diagram displaying the conversion of the retinoblastoma-causing premature stop codon (PTC) to a tryptophan. PAM, ABE editing window, and targeted adenosine are highlighted in blue, red, and green, respectively. (C, D) Frequencies (%) of adenine base editing on gDNA (C) and mRNA (D) in R552X cell lines measured by targeted deep sequencing conducted across four biologically independent experiments (n = 4; mean ± SEM). P values were obtained using one-way analysis of variance (ANOVA) and Tukey’s test for post hoc analysis: *P < 0.05, **P < 0.01, ***P < 0.001.

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