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. 2022 Jul 29;23(15):8416. doi: 10.3390/ijms23158416

Figure 4.

Figure 4

ABE editing in FANCA c.295C > T patient-derived LCL. (A) Editing scheme; In FA patient’s with the Spanish founder mutation, a pmSTOP codon (TAG) is mutated from the WT glutamine (CAG). ABE changes this pmSTOP codon (A > G) to tryptophan (TTG) resulting in translation of the FANCA gene. (B) Alignment of amino acids surrounding the FANCA c.295 region on exon 4 showing variability of the codon in questions across species. (C) B2M Ex.1 SD sgRNA region chromatograms of Sanger sequencing after PCR amplification of B2M exon 1 using EditR. (D) Percentage of B2M editing events (http://baseeditr.com/, accessed on 1 January 2018–1 July 2022) identified by Sanger sequencing 5 days post electroporation. Data are represented as mean ± SD with n = 2 and n = 1 replicates, respectively. (E) FANCA sgRNA region chromatograms of Sanger sequencing after PCR amplification of FANCA exon 4. Reverse complement primer sequence is shown. (F) Percentage of FANCA editing events identified by Sanger sequencing 5 days after electroporation. Data are represented as mean ± SD with n = 2 and n = 1 replicates, respectively.