Fig. 5

GH6-1 knockout by TALEN-mediated genome editing affected Ciona larval development. A, B Control larvae developed from dechorionated eggs. C, D Control larvae developed from eggs electroporated with mVenus plasmids. E, F Control larvae developed from eggs electroporated with single-sided TALEN. G–J Experimental larvae developed from eggs electroporated with paired TALEN plasmids. While most control larvae showed adhesive papillae (p in B–H) and regionally reduced cellulose (arrows in B′–F′), many larvae of GH6-1 TALEN knockout failed to form adhesive papillae (asterisk in J) and show a strong cellulose signal all over the anterior epidermis (J′). K, L Percentages of larvae of each phenotype. K Larvae were grouped as: 3 or 2 papillae, 1 papilla, no papilla. L Cellulose normal: surface cellulose was found at the larval tunic outside the epidermis, while cellulose signal was reduced around papillae in the anterior trunk. Cellulose abnormal: surface cellulose exists, but there is no local reduction of cellulose signal strength around papillae. A–J Are DIC images; B′, D′, F′, H′ and J′ are green fluorescence channels showing cellulose after CBM-GFP staining. c-De, control-dechorionated group. c-Ve, control-mVenus group. L.o, Only the left TALEN plasmid was introduced during electroporation. LR: both of the left and right TALEN plasmids were introduced during electroporation. p: adhesive papillae. Asterisk sign (*): no papilla formation