caa a22 4500 445861355 CHVBK 20180317145444.0 cr unu---uuuuu 170323e20110101xx s 000 0 eng 10.1007/s00439-010-0902-8 doi (NATIONALLICENCE)springer-10.1007/s00439-010-0902-8 Mutation analysis in Bardet-Biedl syndrome by DNA pooling and massively parallel resequencing in 105 individuals [Elektronische Daten] [Sabine Janssen, Gokul Ramaswami, Erica Davis, Toby Hurd, Rannar Airik, Jennifer Kasanuki, Lauren Van Der Kraak, Susan Allen, Philip Beales, Nicholas Katsanis, Edgar Otto, Friedhelm Hildebrandt] Bardet-Biedl syndrome (BBS) is a rare, primarily autosomal-recessive ciliopathy. The phenotype of this pleiotropic disease includes retinitis pigmentosa, postaxial polydactyly, truncal obesity, learning disabilities, hypogonadism and renal anomalies, among others. To date, mutations in 15 genes (BBS1-BBS14, SDCCAG8) have been described to cause BBS. The broad genetic locus heterogeneity renders mutation screening time-consuming and expensive. We applied a strategy of DNA pooling and subsequent massively parallel resequencing (MPR) to screen individuals affected with BBS from 105 families for mutations in 12 known BBS genes. DNA was pooled in 5 pools of 21 individuals each. All 132 coding exons of BBS1-BBS12 were amplified by conventional PCR. Subsequent MPR was performed on an Illumina Genome Analyzer II™ platform. Following mutation identification, the mutation carrier was assigned by CEL I endonuclease heteroduplex screening and confirmed by Sanger sequencing. In 29 out of 105 individuals (28%), both mutated alleles were identified in 10 different BBS genes. A total of 35 different disease-causing mutations were confirmed, of which 18 mutations were novel. In 12 additional families, a total of 12 different single heterozygous changes of uncertain pathogenicity were found. Thus, DNA pooling combined with MPR offers a valuable strategy for mutation analysis of large patient cohorts, especially in genetically heterogeneous diseases such as BBS. Springer-Verlag, 2010 Janssen Sabine Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Ramaswami Gokul Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Davis Erica Center for Human Disease Modeling, Duke University Medical Center, Durham, NC, USA aut Hurd Toby Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Airik Rannar Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Kasanuki Jennifer Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Van Der Kraak Lauren Department of Genetics and Genome Biology, The Hospital for Sick Children, Toronto, Ontario, Canada aut Allen Susan Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Beales Philip Molecular Medicine Unit, UCL Institute of Child Health, London, UK aut Katsanis Nicholas Center for Human Disease Modeling, Duke University Medical Center, Durham, NC, USA aut Otto Edgar Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Hildebrandt Friedhelm Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut Human Genetics Springer-Verlag 129/1(2011-01-01), 79-90 0340-6717 129:1<79 2011 129 439 https://doi.org/10.1007/s00439-010-0902-8 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00439-010-0902-8 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Janssen Sabine Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Ramaswami Gokul Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Davis Erica Center for Human Disease Modeling, Duke University Medical Center, Durham, NC, USA aut NATIONALLICENCE 700 1- Hurd Toby Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Airik Rannar Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Kasanuki Jennifer Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Van Der Kraak Lauren Department of Genetics and Genome Biology, The Hospital for Sick Children, Toronto, Ontario, Canada aut NATIONALLICENCE 700 1- Allen Susan Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Beales Philip Molecular Medicine Unit, UCL Institute of Child Health, London, UK aut NATIONALLICENCE 700 1- Katsanis Nicholas Center for Human Disease Modeling, Duke University Medical Center, Durham, NC, USA aut NATIONALLICENCE 700 1- Otto Edgar Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 700 1- Hildebrandt Friedhelm Departments of Pediatrics and of Human Genetics, University of Michigan, Ann Arbor, MI, USA aut NATIONALLICENCE 773 0- Human Genetics Springer-Verlag 129/1(2011-01-01), 79-90 0340-6717 129:1<79 2011 129 439 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer