Figure 2

Figure 2. Evaluation of germline transmission of ErCas12a-mediated knockin with the MMEJ donor at the tyr locus. (A) Representative images of the red fluorescent reporter expression of F₁ embryos obtained from outcross of the three tyr MMEJ knockin-positive F₀ adults listed in Table 1. Red fluorescence can be observed in the eyes and pigment cells of the embryos. Scale bar: 200 μm. (B) Junction PCR results of the tyr knockin F₁ embryos showing red fluorescence from F₀ #3. Primers tyr F1 and tdt R were used to amplify the 5′ junction, and primers bF1 and tyr R1 were used to amplify the 3′ junction. (C) Sequencing alignment of the junction PCR products from (B) after TA cloning. Black characters indicate sequence of the tyr WT allele, blue characters represent the homologous sequence designed in the donor, and red characters represent the donor sequence.