Medium-scale study of imprinted expression in eight adult tissues of the mouse

The topic of this study is the examination of the allelic expression status of transcripts in mouse. In this approach known and novel transcripts were assessed for their tissue-wide allelic expression status. This is a novel approach as most studies so far only focused on one or few tissues. Besides known imprinted candidates, additional non-coding candidates were predicted from an analysis of published RNA sequencing data. The experimental setting used in this analysis was based on an interspecies cross between two mouse strains, Mus musculus castaneus and Mus musculus FVB that show 20 million SNPs between their genomes. cDNA from different mouse tissues was prepared by RNA isolation from dissected mouse organs and subsequent reverse transcription. A program was devised to allow for the automated design of primers taking into account that the product had to overlap at least 2 SNPs. Purified PCR products were then analysed using high throughput sequencing. The allelic expression of each transcript was determined using Allelome.PRO, a published bioinformatics pipeline.