Complete exon sequencing of all known Usher syndrome genes greatly improves molecular diagnosis

<p>Abstract</p> <p>Background</p> <p>Usher syndrome (USH) combines sensorineural deafness with blindness. It is inherited in an autosomal recessive mode. Early diagnosis is critical for adapted educational and patient management choices, and for genetic counseling. To date, nine causative genes have been identified for the three clinical subtypes (USH1, USH2 and USH3). Current diagnostic strategies make use of a genotyping microarray that is based on the previously reported mutations. The purpose of this study was to design a more accurate molecular diagnosis tool.</p> <p>Methods</p> <p>We sequenced the 366 coding exons and flanking regions of the nine known USH genes, in 54 USH patients (27 USH1, 21 USH2 and 6 USH3).</p> <p>Results</p> <p>Biallelic mutations were detected in 39 patients (72%) and monoallelic mutations in an additional 10 patients (18.5%). In addition to biallelic mutations in one of the USH genes, presumably pathogenic mutations in another USH gene were detected in seven patients (13%), and another patient carried monoallelic mutations in three different USH genes. Notably, none of the USH3 patients carried detectable mutations in the only known USH3 gene, whereas they all carried mutations in USH2 genes. Most importantly, the currently used microarray would have detected only 30 of the 81 different mutations that we found, of which 39 (48%) were novel.</p> <p>Conclusions</p> <p>Based on these results, complete exon sequencing of the currently known USH genes stands as a definite improvement for molecular diagnosis of this disease, which is of utmost importance in the perspective of gene therapy.</p

Comprehensive genetic dissection of wood properties in a widely-grown tropical tree: Eucalyptus

Background: Eucalyptus is an important genus in industrial plantations throughout the world and is grown for use as timber, pulp, paper and charcoal. Several breeding programmes have been launched worldwide to concomitantly improve growth performance and wood properties (WPs). In this study, an interspecific cross between Eucalyptus urophylla and E. grandis was used to identify major genomic regions (Quantitative Trait Loci, QTL) controlling the variability of WPs. Results: Linkage maps were generated for both parent species. A total of 117 QTLs were detected for a series of wood and end-use related traits, including chemical, technological, physical, mechanical and anatomical properties. The QTLs were mainly clustered into five linkage groups. In terms of distribution of QTL effects, our result agrees with the typical L-shape reported in most QTL studies, i.e. most WP QTLs had limited effects and only a few (13) had major effects (phenotypic variance explained &gt; 15%). The co-locations of QTLs for different WPs as well as QTLs and candidate genes are discussed in terms of phenotypic correlations between traits, and of the function of the candidate genes. The major wood property QTL harbours a gene encoding a Cinnamoyl CoA reductase (CCR), a structural enzyme of the monolignol-specific biosynthesis pathway. Conclusions: Given the number of traits analysed, this study provides a comprehensive understanding of the genetic architecture of wood properties in this Eucalyptus full-sib pedigree. At the dawn of Eucalyptus genome sequence, it will provide a framework to identify the nature of genes underlying these important quantitative traits. (Résumé d'auteur

Variants in CUL4B are Associated with Cerebral Malformations

Variants in cullin 4B (CUL4B) are a known cause of syndromic X-linked intellectual disability. Here, we describe an additional 25 patients from 11 families with variants in CUL4B. We identified nine different novel variants in these families and confirmed the pathogenicity of all nontruncating variants. Neuroimaging data, available for 15 patients, showed the presence of cerebral malformations in ten patients. The cerebral anomalies comprised malformations of cortical development (MCD), ventriculomegaly, and diminished white matter volume. The phenotypic heterogeneity of the cerebral malformations might result from the involvement of CUL-4B in various cellular pathways essential for normal brain development. Accordingly, we show that CUL-4B interacts with WDR62, a protein in which variants were previously identified in patients with microcephaly and a wide range of MCD. This interaction might contribute to the development of cerebral malformations in patients with variants in CUL4B

Notable contribution of large CFTR gene rearrangements to the diagnosis of cystic fibrosis in fetuses with bowel anomalies

Grade III fetal bowel hyperechogenicity and/or loop dilatation observed at the second trimester of pregnancy can be due to several disease conditions, including cystic fibrosis (CF). Screening for frequent CF mutations is performed as a first step and, in certain situations, such as when a frequent CF mutation is found in the fetus, the increased risk of CF justifies an in-depth study of the second allele. To determine the contribution of large CFTR gene rearrangements in such cases, detected using a semiquantitative fluorescent multiplex PCR (QFM-PCR) assay, we collated data on 669 referrals related to suspicion of CF in fetuses from 1998 to 2009. Deletions were found in 5/70 cases in which QFM-PCR was applied, dele19, dele22_23, dele2_6b, dele14b_15 and dele6a_6b, of which the last three remain undescribed. In 3/5 cases, hyperechogenicity was associated with dilatation and/or gallbladder anomalies. Of the total cases of CF recognized in the subgroup of first-hand referrals, deletions represent 16.7% of CF alleles. Our study thus strengthens the need to consider large CFTR gene rearrangements in the diagnosis strategy of fetal bowel anomalies, in particular in the presence of multiple anomalies

Confirmation of TFAP2A gene involvement in branchio-oculo-facial syndrome (BOFS) and report of temporal bone anomalies.

International audienceBranchio-oculo-facial syndrome (BOFS) is an autosomal-dominant condition characterized by three main features, respectively: branchial defects, ocular anomalies, and craniofacial defects including cleft lip and/or palate (CL/P). We report on one family with three affected, and two sporadic cases that have been found to carry missense mutations in the newly reported BOFS gene: TFAP2A. This report confirms the involvement of this transcription factor in this developmental syndrome with clinical variability. Moreover, we present CT scan temporal bone anomalies in the familial cases, related to branchial arch defects, highlighting the importance of radiological investigations for differential diagnosis

Un modèle de variabilité fonctionnelle chez les arbres forestiers : le gène CCR d'eucalyptus

National audienceNucleotidic polymorphism of Cinnamoyl CoA Reductase (CCR) gene and its relation with lignin content is studied within a breeding population of Eucalyptus urophylla S.T. Blake (“Timor Mountain Gum”). The nearly full sequence (94%) are obtained for 15 parental trees. This gene (3220 bp) is highly polymorphic showing 131 single nucleotide polymorphism (SNP), 17 insertion-deletions (INDEL), 1 polyA sequence and a microsatellite site. Exons fragments encompass 10 non-synonymous SNPs, half of them within exon 5 (194 bp). Fifteen different haplotypes are reconstructed based on the polymorphism of exon 4 and intron 4. CCR promoting sequence (694 bp) including all the known regulatory sequences is described for the two alleles of one of the genitor trees displaying QTL and CCR gene colocalization in its genetic map. Five SNPs are present. Functional variability of the promoting sequence will be studied in planta through genetic modification of Arabidopsis thaliana. Lignin content was assessed within a sample of 35 full sib families (348 individuals) generated with the 15 parental trees, showing a high genetic additive control for this trait (h²=0.76). A new algorithm based on Reversible-jump MCMC was developed in order to implement association studies. Half of the progeny trees (208) were genotyped using the microsatellite fragment. The results show that a significant part of the observed genetic variance of lignin content is due to the nucleotide polymorphism of the studied gene. Those preliminary results look promising in order to develop early gene assisted selection for eucalyptus clones used as raw material in charcoal and paper production.La variabilité nucléotidique du gène codant la Cinnamoyl CoA Reductase (CCR) et ses effets sur le taux de lignine est étudiée au sein d’une population d’Eucalyptus urophylla S.T. Blake. La presque totalité de la séquence (94%, 3220 paires de bases) est décrite pour 15 individus. Le gène est hautement polymorphe et présente 131 mutations ponctuelles (SNP) ainsi que divers autres types de mutations. Les fragments exoniques présentent 10 SNP non synonymes dont 5 dans l’exon 5. La séquence promotrice (694 pb) est décrite pour les deux allèles d’un des géniteurs. Elle regroupe 5 SNPs. La variabilité fonctionnelle de ce promoteur sera étudiée grâce à son expression dans Arabidopsis thaliana. L’analyse de la teneur en lignine de 348 arbres appartenant à 35 familles de pleins frères obtenues avec ces 15 géniteurs montre que ce caractère présente un fort contrôle génétique additif (h²=0.76). Un nouvel algorithme type MCMC a été développé pour procéder aux études d’association sur 208 descendants génotypés grâce à un marqueur microsatellite présent dans le gène CCR. Les résultats montrent qu’une part importante de la variance du taux de lignine est due au polymorphisme du gène CCR. Ces résultats laissent envisager le développement d’une sélection précoce assistée par marqueurs

CCR gene in Eucalyptus: a model of functional variability in forest trees

La variabilité nucléotidique du gène codant la Cinnamoyl CoA Reductase (CCR) et ses effets sur le taux de lignine est étudiée au sein d’une population d’Eucalyptus urophylla S.T. Blake. La presque totalité de la séquence (94%, 3220 paires de bases) est décrite pour 15 individus. Le gène est hautement polymorphe et présente 131 mutations ponctuelles (SNP) ainsi que divers autres types de mutations. Les fragments exoniques présentent 10 SNP non synonymes dont 5 dans l’exon 5. La séquence promotrice (694 pb) est décrite pour les deux allèles d’un des géniteurs. Elle regroupe 5 SNPs. La variabilité fonctionnelle de ce promoteur sera étudiée grâce à son expression dans Arabidopsis thaliana. L’analyse de la teneur en lignine de 348 arbres appartenant à 35 familles de pleins frères obtenues avec ces 15 géniteurs montre que ce caractère présente un fort contrôle génétique additif (h²=0.76). Un nouvel algorithme type MCMC a été développé pour procéder aux études d’association sur 208 descendants génotypés grâce à un marqueur microsatellite présent dans le gène CCR. Les résultats montrent qu’une part importante de la variance du taux de lignine est due au polymorphisme du gène CCR. Ces résultats laissent envisager le développement d’une sélection précoce assistée par marqueurs.Nucleotidic polymorphism of Cinnamoyl CoA Reductase (CCR) gene and its relation with lignin content is studied within a breeding population of Eucalyptus urophylla S.T. Blake (“Timor Mountain Gum”). The nearly full sequence (94%) are obtained for 15 parental trees. This gene (3220 bp) is highly polymorphic showing 131 single nucleotide polymorphism (SNP), 17 insertion-deletions (INDEL), 1 polyA sequence and a microsatellite site. Exons fragments encompass 10 non-synonymous SNPs, half of them within exon 5 (194 bp). Fifteen different haplotypes are reconstructed based on the polymorphism of exon 4 and intron 4. CCR promoting sequence (694 bp) including all the known regulatory sequences is described for the two alleles of one of the genitor trees displaying QTL and CCR gene colocalization in its genetic map. Five SNPs are present. Functional variability of the promoting sequence will be studied in planta through genetic modification of Arabidopsis thaliana. Lignin content was assessed within a sample of 35 full sib families (348 individuals) generated with the 15 parental trees, showing a high genetic additive control for this trait (h²=0.76). A new algorithm based on Reversible-jump MCMC was developed in order to implement association studies. Half of the progeny trees (208) were genotyped using the microsatellite fragment. The results show that a significant part of the observed genetic variance of lignin content is due to the nucleotide polymorphism of the studied gene. Those preliminary results look promising in order to develop early gene assisted selection for eucalyptus clones used as raw material in charcoal and paper production