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Qin et al., 2018 - Precise A•T to G•C base editing in the zebrafish genome. BMC Biology   16:139 Full text @ BMC Biol.

Fig. 1

Comparison of the original ABE system and optimized zABE7.10 system in zebrafish. a Schematic diagrams of two adenine base-editing system, pCMV-ABE and zABE7.10. The zABE7.10 system is a zebrafish codon optimized version by IGE. b Selected sites that can work efficiently in zebrafish using BE system or Target-AID system. Target sequence (black) and PAM region (blue) are indicated. c Sequencing result at the ddx17-g1 target in pCMV-ABE system in zebrafish. d Sequencing result at the ddx17-g1 target in zABE7.10 system in zebrafish

Fig. 2

Codon-optimized zABE7.10 system in zebrafish induces A to G base conversion in zebrafish. a Schematic view of the gRNA target site in rps14 gene and the sequencing results. b Morphological phenotype of rps14E12G/− deficient embryos. co-dianisidine staining results of the rps14E12G/− embryos. d Schematic view of the gRNA target site in musk gene and the sequencing results. e Sequence chromatograms of atp5b locus. f Sequence chromatograms of wu:fc01d11 locus. g Comparison the base editing efficiency between zABE7.10-nickcase Cas9 and zABE7.10-dCas9 system. Target sequence (black), PAM region (blue), and the substituted bases (red) are indicated. The black line presents the changed amino acid. The red arrows indicate the overlapped peaks. The black dashes and blue dashes denote deleted bases and inserted bases in the sequence, respectively. The red asterisks indicated the overlapping peaks outside the targeted A

Fig. 3

Summary of adenine base-editing results in zebrafish. a The base conversion efficiency, cleavage activities, indels frequency, and germline-targeting efficiency of five target sites (ddx17-g1, rps14, atp5b, wu:fc01d11 and musk) are summarized. b, c The sequence chromatograms results of zABE6.3, zABE7.8, and zABE7.9 in the rps14 and musk target site. Target sequence (black), PAM region (blue), and the substituted bases (red) are indicated. The red arrows indicate the overlapped peaks

Fig. 4

Adenine base editors modified by bpNLS enhance the base editing efficiency in zebrafish. a Schematic diagrams of three adenine base-editing system, zABE7.10, zABE7.10max, and zABE7.10-GE. The zABE7.10 system is a zebrafish codon-optimized version by IGE with SV40 NLS. The zABE7.10max is a version with bis-bpNLS and IGE codons. zABE7.10-GE system contain a GenScript-codon optimized sequence and bis-bpNLS. b The comparison of base editing efficiency of three versions of adenine base editor in zebrafish. c, f The sequencing results of the rps14, atp5b, wu:fc01d11, and musk gene in the zABE7.10max system. Target sequence (black), PAM region (blue), and the substituted bases (red) are indicated. The red arrows indicate the overlapped peaks. Values and error bars represent the mean of n = 3 biologically independent experiments. *p < 0.05

Acknowledgments:
ZFIN wishes to thank the journal BMC Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ BMC Biol.