Non-exomic and synonymous variants in ABCA4 are an important cause of Stargardt disease.
Non-exomic and synonymous variants in ABCA4 are an important cause of Stargardt disease.
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DOI：
10.1093/hmg/ddt367
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发表时间：
2013-12-20
影响因子：
3.5
通讯作者：
Stone EM
中科院分区：
文献类型：
--
作者：
Braun TA;Mullins RF;Wagner AH;Andorf JL;Johnston RM;Bakall BB;Deluca AP;Fishman GA;Lam BL;Weleber RG;Cideciyan AV;Jacobson SG;Sheffield VC;Tucker BA;Stone EM
ABCA4突变导致Stargardt病和其他致盲的常染色体隐性视网膜疾病。然而，对具有Stargardt病临床特征的患者的完整编码序列进行测序有时无法检测到一种或两种突变。例如，在208个个体中，有明确的临床证据的ABCA4疾病在一个单一的机构确定，28个只有一个致病等位基因中确定的外显子和剪接点的主要视网膜转录的基因。对这28个先证者进行单倍型分析，发现10个家系共有3个单倍型，这表明28个缺失等位基因中有18个是罕见的，仅在队列中出现一次。我们假设ABCA 4中罕见的替代剪接点附近的突变可能会通过增加这些位点错误剪接的可能性而导致疾病。从人类供体眼睛中提取的RNA的下一代测序揭示了十几个交替外显子，这些外显子偶尔会并入正常人视网膜中的ABCA 4转录本中。我们对28名单等位基因受试者的基因组DNA进行了测序，发现在外显子36.1的剪接信号中存在两种不同的变异，这在正常个体中是不存在的（P < 10−6）。从具有这些突变的患者的角质形成细胞获得的RNA的分析揭示了预测的替代转录物。这项研究说明了人类供体组织和患者来源的细胞系的RNA序列分析的实用性，以确定外显子组测序检测不到的突变。
Mutations in ABCA4 cause Stargardt disease and other blinding autosomal recessive retinal disorders. However, sequencing of the complete coding sequence in patients with clinical features of Stargardt disease sometimes fails to detect one or both mutations. For example, among 208 individuals with clear clinical evidence of ABCA4 disease ascertained at a single institution, 28 had only one disease-causing allele identified in the exons and splice junctions of the primary retinal transcript of the gene. Haplotype analysis of these 28 probands revealed 3 haplotypes shared among ten families, suggesting that 18 of the 28 missing alleles were rare enough to be present only once in the cohort. We hypothesized that mutations near rare alternate splice junctions in ABCA4 might cause disease by increasing the probability of mis-splicing at these sites. Next-generation sequencing of RNA extracted from human donor eyes revealed more than a dozen alternate exons that are occasionally incorporated into the ABCA4 transcript in normal human retina. We sequenced the genomic DNA containing 15 of these minor exons in the 28 one-allele subjects and observed five instances of two different variations in the splice signals of exon 36.1 that were not present in normal individuals (P < 10−6). Analysis of RNA obtained from the keratinocytes of patients with these mutations revealed the predicted alternate transcript. This study illustrates the utility of RNA sequence analysis of human donor tissue and patient-derived cell lines to identify mutations that would be undetectable by exome sequencing.
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来自1,092个人基因组的遗传变异的综合图。
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
4.4
作者：
Zernant, Jana;Schubert, Carl;Allikmets, Rando
通讯作者：
Allikmets, Rando
影响因子：
3.5
作者：
Schindler EI;Nylen EL;Ko AC;Affatigato LM;Heggen AC;Wang K;Sheffield VC;Stone EM
通讯作者：
Stone EM
影响因子：
--
作者：
Downs, Katy;Zacks, David N.;Ayyagari, Radha
通讯作者：
Ayyagari, Radha
影响因子：
3.5
作者：
Cideciyan, Artur V.;Swider, Malgorzata;Palczewski, Krzysztof
通讯作者：
Palczewski, Krzysztof