ZFIN Image: Moreno-Mateos et al., 2017, Fig. S5

Image

Figure Caption

Fig. S5

LbCpf1-crRNA RNP complexes are an efficient genome editing system in X. tropicalis.

a. Diagram illustrating 3 crRNAs (orange) targeting slc45a2 and tyr exon 1 in X. tropicalis.

b. Phenotypes obtained after injection of the AsCpf1-crRNA or LbCpf1-crRNA RNP complexes (20 fmol) containing a mix of 3 crRNAs targeting slc45a2 (alb) or tyr in X. tropicalis. Lateral views (top; scale bar, 1 mm) and insets of the eyes (bottom; scale bar, 0.4 mm) of stage 47–48 embryos are shown.

c. Phenotypic evaluation of Cpf1-crRNA RNP complexes (20 fmol) injections targeting slc45a2 (alb) or tyr in X. tropicalis. Stacked barplots showing the percentage of severe mutant (light gray), mild mutant (dark grey) and phenotypically WT (black) embryos at stage 47-48. Number of embryos evaluated (n) is shown for each condition.

d. In vitro cleavage assay using 30 pmol of AsCpf1 RNP or LbCpf1 RNP complexes containing a mix of 3 crRNAs (Supplementary Fig. 5a) and a ~380pb (~ 0.45 pmol) or ~440pb (~ 0.38 pmol) PCR product including crRNA target sites for slc45a2 (alb) (left) and tyr (right), respectively. Incubations were carried out at 37°C and 25°C for 90 min. MW: Molecular weight marker (kilobase).

Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.