ZFIN Image: Liang et al., 2022, Fig. 4

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Figure Caption

Fig. 4

Efficient adenine base editing mediated by zSpRY-ABE8e across various PAMs.

a The mRNA construct of zSpRY-ABE8e used for adenine base editing in zebrafish. b Summary of A-to-G base editing efficiency of various loci with NRN PAMs induced by zSpRY-ABE8e editor in zebrafish. The position of editing base in the gRNA was labelled with numbers. (Values are presented as mean value ± standard deviation (SD), n = 3 biological replicates.) c Schematic diagram and sequencing results of tsr2 (M1V) mutation induced by zSpRY-ABE8e in zebrafish. The PAM sequences are underlined in red, the detected nucleotide changes are highlighted in blue, the targeted amino acids are highlighted in bold, and the nucleotide substitutions are indicated with a red arrowhead in the sequencing chromatograms. d Morphological phenotype of tsr2 ^(M1V)/⁻-deficient embryos at 2 dpf. The comparison of eye size is underlined with a red frame in the lateral view and with diameter measurement in the dorsal view. Red arrows indicate a pointy head and slight pericardial oedema of tsr2 ^(M1V)/− mutants. Scale bar: 500 μm. This experiment was repeated three times independently with similar results. e On-target, product purity and off-target analysis of zSpRY-ABE8e induced A-to-G editing at rpl17-NTA, rpl9-NGT and tsr2-NGT sites using NGS. The PAM sequences are underlined in red. NA: Not Applicable. All source data in this figure are provided as a Source data file.

Acknowledgments

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