FIGURE

Fig. 6

ID

ZDB-FIG-131031-19

Publication

Jao et al., 2013 - Efficient multiplex biallelic zebrafish genome editing using a CRISPR nuclease system

Other Figures

All Figure Page

Back to All Figure Page

Fig. 6

Multiplex genome editing in zebrafish by Cas9. A mix of five gRNAs (penta gRNAs: ddx19 gRNA, 55 pg; egfp gRNA, 15 pg; gol gRNA, 25 pg; mitfa gRNA, 60 pg; and tyr gRNA, 25 pg) were coinjected with nls-zCas9-nls RNA (150 pg) into Tg(-5.1mnx1:egfp) transgenic embryos. (A–E) Lateral views of the wild-type (A–A22) and the penta gRNAs/nls-zCas9-nls RNA-injected (B–B22 and C–E) embryos. The combination of three distinct phenotypes—brain necrosis/small eyes/curved body axis, hypopigmentation, and loss of EGFP expression in the motoneurons—caused by biallelic inactivations of multiple target genes simultaneously were observed in the same embryos (B–B22 and E and E2). Some quintuple knockout embryos showed no EGFP-positive motoneurons due to the almost nonmosaic disruption of the Tg(-5.1mnx1:egfp) transgene, whose presence was confirmed by PCR (E2). (F) T7EI assays showed high mutagenesis rates at five genomic targets in six randomly selected quintuple knockout embryos (penta KO 1–6: D, ddx19; E, egfp; G, gol; M, mitfa; T, tyr). (Scale bars: in A, 300 μm; in B22, 50 μm; in E, 500 μm.)

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

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 @ Proc. Natl. Acad. Sci. USA