Horb M , Wlizla M , Abu-Daya A , McNamara S , Gajdasik D , Igawa T , Suzuki A , Ogino H , Noble A , Centre de Ressource Biologique Xenope team in France , Robert J , James-Zorn C , Guille M .

P41 HD064556 NICHD NIH HHS , R24 AI059830 NIAID NIH HHS , Wellcome Trust , P40 OD010997 NIH HHS , R21 AI139718 NIAID NIH HHS , NC/P001009/1 National Centre for the Replacement, Refinement and Reduction of Animals in Research

FIGURE 1. Labeling haematopoietic stem cells in live embryos. Xenopus laevis eggs were microinjected with CRISPR/Cas9 targeting the 3′UTR of runx1 together with a DNA construct containing 400 bp homology arms from either side of the cut site flanking an IRES controlling eGFP expression. Founder embryos were screened for mosaic expression of GFP in the correct regions and 24 grown to adulthood. Their offspring were then screened for germline transmission; two sets of offspring showed strong expression in the vbi (A, arrowed) and vasculature (B, examples arrowed) as expected for runx1 at this stage. The transmission rate was 46 and 52% in the 2 sets. An F2 embryo is shown.

FIGURE 2. Genetic relationships and heterozygosity reduction curve in the strains of X. tropicalis. (A) Neighbor joining trees of the strains based on genetic distances (DA) of 60 Simple Sequence Length Polymorphism (SSLP) markers (upper) and p-distance of mitochondrial haplotypes (2,328 bp; lower). Numbers on branches indicate percent bootstrap probability (only indicated > 50%). (B) Plot of expected heterozygosity (HE) against generation number (t). The curved dotted strain indicates theoretical reduction of heterozygosity via single brother-sister matings for every generation, calculated from the following equation: HE = H0(1−12Ne)t.

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