Combining GWAS and RNA-Seq Approaches for Detection of the Causal Mutation for Hereditary Junctional Epidermolysis Bullosa in XXXX .

@entity1 @entity71839 @entity742 @entity2640 @entity3202 @entity67617 @entity74 @entity2321
UNASSIGNED: In this study, we demonstrate the use of a genome-wide association mapping together with RNA-seq in a reduced number of samples, as an efficient approach to detect the causal mutation for a @entity74 . @entity3202 is a recessive genodermatosis that manifests with neonatal mechanical fragility of the skin, blistering confined to the lamina lucida of the basement membrane and severe alteration of the hemidesmosomal junctions. In Spanish Churra @entity742 , @entity3202 ( @entity3202 ) has been detected in two commercial flocks. The @entity3202 locus was mapped to @entity742 chromosome 11 by GWAS and subsequently fine-mapped to an 868-kb homozygous segment using the identical-by-descent method. The @entity67617 , which is located within this region, was identified as the best positional and functional candidate gene. The RNA-seq variant analysis enabled us to discover a 4-bp deletion within exon 33 of the @entity67617 gene ( @entity71839 ). The @entity71839 mutation causes a frameshift resulting in a premature stop codon at position 1472 of the integrin b4 protein. A functional analysis of this deletion revealed decreased levels of mRNA in @entity3202 skin samples and the absence of integrin b4 labeling in immunohistochemical assays. Genotyping of @entity71839 showed perfect concordance with the recessive mode of the disease phenotype. Selection against this causal mutation will now be used to solve the problem of @entity3202 in flocks of Churra @entity742 . Furthermore, the identification of the @entity67617 mutation means that affected @entity742 can be used as a large @entity1 animal model for the @entity1 form of @entity2640 with @entity2321 . Our approach evidences that RNA-seq offers cost-effective alternative to identify variants in the species in which high resolution exome-sequencing is not straightforward.
