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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. |