In addition, when no other mutation outside the original family was found, functional studies as well

as modelling in the animals were performed. Table 2. Muscle disease gene discovery by NGS. The first example is the targeted NGS of 640 exons from a chromosomal region located on chromosome 5q23, identified by phased haplotype analysis that was used to discover the cause of EMARDD, a disease characterized by early onset myopathy, areflexia, respiratory distress and dysphagia (17). These infantile myopathies Inhibitors,research,lifescience,medical with diaphragmatic paralysis are genetically heterogeneous and clinical symptoms do not assist in differentiating between them. EMARDD is inherited as an autosomal recessive disorder. Affected member of a consanguineous family from Pakistan showed a homozygous 10-bp duplication (c.2288_2297dup) in the coding Inhibitors,research,lifescience,medical sequence of exon 19 of MEGF10 (multiple epidermal growth factor-like domains-10 protein). Other independent families

were homozygous or compound heterozygous for other lossof- function mutations in MEGF10, thus proving proof of the causative role for this Inhibitors,research,lifescience,medical gene. MEGF10 is a regulator of satellite cell myogenesis, highly expressed in activated satellite cells, that regulates their proliferation, differentiation, and fusion into multinucleated myofibers, which are greatly reduced in muscle. A second example is the identification of the cause of a form of congenital myopathy with prominent internal Sorafenib ic50 nuclei and atypical cores (18). Congenital myopathies are well suited for whole exome NGS since they are clinically and genetically heterogeneous diseases. In this case the Authors performed a SNP linkage analysis on ten Inhibitors,research,lifescience,medical individuals (including five affected members) of a family with autosomal dominant inheritance characterized by distal weakness and corelike areas and increased internalized nuclei at biopsy. The top LOD score was only 1.87 on chromosome 16. The DNA from the index case alone was analyzed by whole-exome sequencing using the

Inhibitors,research,lifescience,medical NimbleGen exome capture and NGS. Among many unique variants, the disease was linked to a heterozygous C>T change at c.68-1 of CCDC78, an uncharacterized coiled-coiled domain-containing gene located on 16p13 and expressed in skeletal muscle. This change alters the splicing of exon 2. The mutation was confirmed TCL in the original family and tested in the zebrafish using a morpholino- mediated splice-site alteration. The CCDC78 alteration in zebrafish resulted in altered motor function and abnormal muscle ultrastructure. A third example is the use of whole-exome NGS or traditional positional cloning by two different groups to reveal the causative gene in an autosomal dominant limb-girdle muscular dystrophy (LGMD1D). LGMD1D is characterized by skeletal muscle vacuoles, previously mapped to chromosome 7q36. Sarparanta et al. performed the characterization of LGMD1D in Finnish families and refined the locus to a 3.4-Mb region containing 12 genes.