Fig 1

The self-cleaving T3H48 ribozyme induces pre-mRNA cleavage in zebrafish.

(a) Schematic of the dual ubb promoter vector that controls the expression of an eGFP containing a human β-globin (hHBB) intron and mCherry; the active and inactive ribozymes are inserted in the hHBB intron; red x indicates the location of the ribozyme inactivating mutation. (b) Activity of the N79, N107, N117, T3H48, and 3xT3H48 ribozymes represented as the ratio of eGFP mRNA levels in embryos injected with the active or inactive ribozyme construct; mCherry was used as a reference gene in this experiment to account for the variable number of plasmid copies in each injected embryo; n=3 biologically independent samples; Ct values are listed in S1 Table. (c-d) Merge of brightfield and fluorescence images of 25 hpf (hours post-fertilization) embryos injected at the one-cell stage with the dual ubb vector, comprising an inactive (c) or active (d) T3H48-HHR. The proportion of embryos matching the image shown is indicated in the top right corner of each image. The diagram indicates the Anterior-Posterior (A-P) and Dorsal-Ventral (D-V) axes.