ZFIN Image: Ablain et al., 2015, Fig. S3

Image

Figure Caption

Fig. S3

Fluorescent phenotype induced by CRISPR vectors targeting urod (related to Figure 3).

(A) Confocal images of 50 hpf embryos injected with Tol2 mRNA and pcmlc2:GFP, U6:gRNA urod vectors expressing Cas9 under the control of the indicated promoters.

(B) Quantification of the fluorescent phenotype presented in Figure 3A. WT: no fluorescent blood cell. Mild: <10 fluorescent blood cells. Intermediate: <50 fluorescent blood cells. Strong: >50 fluorescent blood cells. Ubi:Cas9 (n=61), gata1:Cas9 (n=51), mylz2:Cas9 (n=43).

(C) Schematic representation of a variant tissue-specific CRISPR vector. Gata1 promoter (Pgata1) drives both Cas9 and GFP expression through a self-cleaving T2A peptide. Any gRNA of interest can be produced ubiquitously off the U6 promoter. Tol2 indicates transposition sites for the Tol2 transposase. pA: SV40 polyA sequence.

(D) Confocal images of the yolk region of 30 hpf embryos injected with Tol2 mRNA and the pA2, U6:gRNA urod, gata1:Cas9-T2A-GFP vector. Arrows point to cells that are both green and red. Scale bar: 20 μm. Note the small size of red-only cells.

Acknowledgments

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Reprinted from Developmental Cell, 32(6), Ablain, J., Durand, E.M., Yang, S., Zhou, Y., Zon, L.I., A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in Zebrafish, 756-64, Copyright (2015) with permission from Elsevier. Full text @ Dev. Cell