High-throughput, pooled sequencing identifies mutations in NUBPL and FOXRED1 in human complex I deficiency
High-throughput, pooled sequencing identifies mutations in NUBPL and FOXRED1 in human complex I deficiency
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DOI：
10.1038/ng.659
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发表时间：
2010-10-01
期刊：
影响因子：
30.8
通讯作者：
Mootha, Vamsi K.
中科院分区：
文献类型：
--
作者：
Calvo, Sarah E.;Tucker, Elena J.;Mootha, Vamsi K.
发现线粒体呼吸链疾病的分子基础是具有挑战性的，因为涉及大量的线粒体和核基因。我们报告了一个集中的候选基因预测，高通量测序和实验验证的策略，以揭示线粒体复合物I疾病的分子基础。我们从103例病例和42例健康对照组中创建了7个DNA池，然后对103个候选基因进行了深度测序，以确定151个预计会影响蛋白质功能的罕见变异。我们在60个以前未解决的病例中的13个中建立了基因诊断，使用验证性实验，包括cDNA互补，以表明NUBPL和FOXRED 1的突变可以导致复合物I缺乏。我们的研究说明了如何大规模测序，加上功能预测和实验验证，可以用来确定个别情况下的因果突变。
Discovering the molecular basis of mitochondrial respiratory chain disease is challenging given the large number of both mitochondrial and nuclear genes that are involved. We report a strategy of focused candidate gene prediction, high-throughput sequencing and experimental validation to uncover the molecular basis of mitochondrial complex I disorders. We created seven pools of DNA from a cohort of 103 cases and 42 healthy controls and then performed deep sequencing of 103 candidate genes to identify 151 rare variants that were predicted to affect protein function. We established genetic diagnoses in 13 of 60 previously unsolved cases using confirmatory experiments, including cDNA complementation to show that mutations in NUBPL and FOXRED1 can cause complex I deficiency. Our study illustrates how large-scale sequencing, coupled with functional prediction and experimental validation, can be used to identify causal mutations in individual cases.