Fig. 2

PreMA TALEN reagent can be used to recapitulate previously reported loss-of-chrd-function phenotype in 1 dpf F0, injected larvae.

A. Top–Wildtype chrd sequence with TALEN binding sites annotated in teal. The dotted red boxes are MH arms predicted to be used most frequently. Raw sequence alignment of the whole PCR amplicon demonstrates that the majority of reads are the expected 7 bp deletion allele. Bottom–summary data from subcloning analyses. 50% of the mutant allele recovered were of the predicted MH allele. B. Previously reported chrd loss-of-function phenotype was successfully recapitulated using this TALEN pair. Phenotype severity was graded by the degree of Intermediate-Cell-Mass expansion in the tail and by the reduced head size by 1 dpf. Box plot demonstrating phenotypic penetrance is provided with each experiment denoted by a unique marker shape. N = 3 biological and technical replicates. At least 29 injected animals were scored in each experiment.