FIGURE

Fig. 3

ID

ZDB-FIG-220617-35

Publication

Liang et al., 2022 - SpG and SpRY variants expand the CRISPR toolbox for genome editing in zebrafish

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Fig. 3

Efficient cytosine base editing mediated by SpRY-CBE4max across various PAMs.

a The mRNA construct of SpRY-CBE4max used for cytosine base editing in zebrafish. b Summary of C-to-T base editing efficiency of various loci with NRN PAMs induced by SpRY-CBE4max editor in zebrafish. The position of the 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 mitfa (E318K) mutation induced by SpRY-CBE4max. The PAM sequences are underlined in red, the detected nucleotide changes are highlighted in blue, the targeted amino acids are highlighted in bold, the nucleotide substitutions are indicated by a red arrowhead in the sequencing chromatograms, and the synonymous mutations induced by SpRY-CBE4max are indicated with a blue arrowhead. d Lateral view of 2 dpf F0 embryos injected with mitfa (E318K) gRNA and SpRY-CBE4max mRNA showing pigmentation defects. Scale bar: 1 mm. This experiment was repeated three times independently with similar results. e On-target, product purity and off-target analysis of SpRY-CBE4max induced C-to-T editing at rpl17-NTA, ddx21-NGA and mitfa-NAT 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.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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