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Fig. 1
Motor neuron (MN) specific expression of KillerRed (KR). (A) A membrane localization signal (MLS) targets the photosensitizer protein KillerRed (KR) to the intracellular cell membrane of MNs (mnx1 promoter). Upon green light illumination (LI,), KR induces lipid oxidization generating reactive oxygen species (ROS) alongside photo-bleaching of KR. (B) Synthetic transposase mRNA and a Tol2 transposon plasmid DNA construct containing the Tol2 element, the mnx1 promoter and the sequence encoding MLS-KR were co-injected into one cell stage zebrafish eggs. The Tol2 construct is excised from the plasmid DNA and integrated into the genomic DNA. Tol2 insertions in germ cells are transmitted to the F1 generation (modified after Kawakami et al., 2007). (C) PCR analysis of genomic DNA extracted from 24hpf F1 generation zebrafish embryos, confirmed germ line transmission of KR. Expected product size for MLS-KR was 531 bp, b-actin served as a positive control (housekeeping gene). (D-E) MN specific MLS-KR expression (red) at 3 dpf (Tg[mnx1:MLS-KillerRed]). Images are lateral views, anterior to the left, dorsal to the top. Scale bar 25 µm.