Fig. 2

Mutation in kri1l gene is responsible for cas002 phenotypes. (A-C) Morpholino knockdown of kri1l phenocopies cas002 mutant. kri1l ATG MO and splicing MO were injected into wild-type embryos at one-cell stage. At 3 dpf, the injected embryos were fixed and analyzed for cmyb expression by WISH. The percentage of morphants with the reduced cmyb expression phenotype is listed at the bottom of B and C. (D-F) Transient expression of wild-type kri1l mRNA rescues cas002 mutant. cas002 mutant embryos at one-cell stage were injected with synthesized wild type kri1l mRNA. The injected embryos were fixed for analysis of cmyb gene expression using WISH at 4 dpf. After WISH and photographing, all embryos were genotyped by sequencing of genomic DNA; the percentage of the rescue was then evaluated. The percentage of fully rescued mutant embryos is about 54% (50/92), while the rest are partially rescued. (A2-F2) Details of cmyb expression in CHT regions in A-F. (G-H) E-Bioanalyser analysis of total RNA isolated from WT and kri1l^cas002 embryo pools (each pool of 12 embryos) at 3 dpf. A significant reduction in the 18S peak but unchanged 28S peak in kri1l^cas002 results in an elevated 28S/18S rRNA ratio. (I) The 18S rRNA reduction can be restored in mutant embryos by injection of kri1l^WT mRNA.