biostudies-literatureCalvo SENIDDK NIH HHSNIGMS NIH HHS851-8https://www.ebi.ac.uk/biostudies/studies/S-EPMC2977978biostudies-literatureUnknown42(10)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 illustraNature geneticsHigh-throughput, pooled sequencing identifies mutations in NUBPL and FOXRED1 in human complex I deficiency.PMC2977978GM077465R01 GM077465P30 DK043351Altshuler DCalvo SEBurtt NPDaly MJWilson CJRivas MCrawford GTucker EJGoldberger OAWiltshire EKirby DMRedman MCMootha VKGuiducci CGabriel SBCompton AGThorburn DRBruno DLfalseHigh-throughput, pooled sequencing identifies mutations in NUBPL and FOXRED1 in human complex I deficiency.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 illustra2010-01-01T00:00:00Z2010 Oct2026-05-02T18:26:39.496Z2026-04-07T18:19:36.401ZS-EPMC29779782081838310.1038/ng.659