Targeted Capture and Next-Generation Sequencing Identifies C9orf75, Encoding Taperin, as the Mutated Gene in Nonsyndromic Deafness DFNB79
Targeted Capture and Next-Generation Sequencing Identifies C9orf75, Encoding Taperin, as the Mutated Gene in Nonsyndromic Deafness DFNB79
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DOI：
10.1016/j.ajhg.2010.01.030
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发表时间：
2010-03-12
影响因子：
9.8
通讯作者：
Friedman, Thomas B.
中科院分区：
文献类型：
--
作者：
Rehman, Atteeq Ur;Morell, Robert J.;Friedman, Thomas B.
靶向基因组捕获结合下一代测序用于分析染色体9q34.3上的2.9 Mb DFNB 79区间，其包括108个候选基因。基因组DNA从一个受影响的成员的血缘家庭分离隐性，非综合征性听力损失被用来制作一个文库的片段覆盖DFNB 79连锁区间定义的遗传分析的四个家系。通过评估402，554个测序读数的文库检测转录序列中8个先前未报道的变体的纯合性，随后通过桑格测序证实。在这些变异体中，有6个在巴基斯坦人群中被确定为多态性，1个在非编码基因中，该基因随后被排除在DFNB 79连锁区间之外。剩下的变异是预测基因C9orf75中的无义突变，改名为TPRN。对其他三个DFNB 79连锁家族的评估鉴定了三个额外的移码突变，总共四个该基因的截短等位基因。尽管TPRN在许多组织中表达，但该蛋白产物在小鼠耳蜗中的免疫定位显示，在毛细胞静纤毛的锥形区域有显着表达。因此，我们将这种蛋白质命名为Taperin。
Targeted genome capture combined with next-generation sequencing was used to analyze 2.9 Mb of the DFNB79 interval on chromosome 9q34.3, which includes 108 candidate genes. Genomic DNA from an affected member of a consanguineous family segregating recessive, nonsyndromic hearing loss was used to make a library of fragments covering the DFNB79 linkage interval defined by genetic analyses of four pedigrees. Homozygosity for eight previously unreported variants in transcribed sequences was detected by evaluating a library of 402,554 sequencing reads and was later confirmed by Sanger sequencing. Of these variants, six were determined to be polymorphisms in the Pakistani population, and one was in a noncoding gene that was subsequently excluded genetically from the DFNB79 linkage interval. The remaining variant was a nonsense mutation in a predicted gene, C9orf75, renamed TPRN. Evaluation of the other three DFNB79-linked families identified three additional frameshift Mutations, for a total of four truncating alleles of this gene. Although TPRN is expressed in many tissues, immunolocalization of the protein product in the mouse cochlea shows prominent expression in the taper region of hair cell stereocilia. Consequently, we named the protein taperin.