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Fig. 3
Mutagenesis of cnrip1a and cnrip1b. (a) Alignments of wild type with each of three mutant alleles showing the predicted expressed mutant polypeptide beneath. Bold underline indicates the gRNA target, red font the protospacer motif recognised by Cas9, hyphens deleted bases, blue highlight inserted bases and asterisks novel stop codons. In the mutant protein sequences, bold text indicates the residual wild type fragment and normal font the aberrant polypeptide tail. (b,c). To test for nonsense-mediated decay, about 50 siblings from in-crosses of cnrip1a +/kg98 (b) and cnrip1b +/kg101 (c) mutant carriers were subjected to in situ hybridisation for the cognate mRNA at the indicated stages, photographed, DNA extracted, PCR performed across the mutant locus and genotype confirmed by DNA sequencing as indicated beneath each panel. Quantitative evidence of reproducibility is given in Table S1.