Recovery of Ccdc151Snbl Mouse Mutation by Whole-Mouse Exome Sequencing Analysis
The pathogenic mutation in the Snowball ENU mutant (b2b1885Clo, MGI 5445347) was identified by whole mouse exome sequence capture performed using the Agilent SureSelect Mouse All Exon V1 kit followed by pair-end sequencing using an Illumina HiSeq 2000 sequencer. Average 51.8× target coverage was achieved. Reads were mapped to the C57BL/6 reference genome (mm9) using CLCBio Genomic Workbench and GATK v.2.8 software. Sequence variants were annotated with Annovar and filtered against dbSNP and our in-house mouse exome databases with custom scripts. Five homozygous coding variants were obtained in the single homozygous Snbl mutant analyzed by exome sequencing analysis. Genotyping for all five coding variants in two additional Snbl mutants exhibiting heterotaxy or situs inversus totalis identified only a c.828+2T>C Ccdc151 variant as consistently homozygous in all of the Snbl mutant offspring. This was further confirmed with additional breeding and genotyping/phenotyping analysis.