Journal article
Exome sequencing and analysis of induced pluripotent stem cells identify the cilia-related gene male germ cell-associated kinase (MAK) as a cause of retinitis pigmentosa
Proceedings of the National Academy of Sciences - PNAS, Vol.108(34), pp.E569-E576
08/23/2011
DOI: 10.1073/pnas.1108918108
PMCID: PMC3161526
PMID: 21825139
Abstract
Retinitis pigmentosa (RP) is a genetically heterogeneous heritable disease characterized by apoptotic death of photoreceptor cells. We used exome sequencing to identify a homozygous Alu insertion in exon 9 of male germ cell-associated kinase (MAK) as the cause of disease in an isolated individual with RP. Screening of 1,798 unrelated RP patients identified 20 additional probands homozygous for this insertion (1.2%). All 21 affected probands are of Jewish ancestry. MAK encodes a kinase involved in the regulation of photoreceptor-connecting cilium length. Immunohistochemistry of human donor tissue revealed that MAK is expressed in the inner segments, cell bodies, and axons of rod and cone photoreceptors. Several isoforms of MAK that result from alternative splicing were identified. Induced pluripotent stem cells were derived from the skin of the proband and a patient with non-MAK-associated RP (RP control). In the RP control individual, we found that a transcript lacking exon 9 was predominant in undifferentiated cells, whereas a transcript bearing exon 9 and a previously unrecognized exon 12 predominated in cells that were differentiated into retinal precursors. However, in the proband with the Alu insertion, the developmental switch to the MAK transcript bearing exons 9 and 12 did not occur. In addition to showing the use of induced pluripotent stem cells to efficiently evaluate the pathogenicity of specific mutations in relatively inaccessible tissues like retina, this study reveals algorithmic and molecular obstacles to the discovery of pathogenic insertions and suggests specific changes in strategy that can be implemented to more fully harness the power of sequencing technologies.
Details
- Title: Subtitle
- Exome sequencing and analysis of induced pluripotent stem cells identify the cilia-related gene male germ cell-associated kinase (MAK) as a cause of retinitis pigmentosa
- Creators
- Budd A Tucker - Department of Ophthalmology and Visual Sciences, University of Iowa Carver College of Medicine, Iowa City, IA 52242, USATodd E ScheetzRobert F MullinsAdam P DeLucaJeremy M HoffmannRebecca M JohnstonSamuel G JacobsonVal C SheffieldEdwin M Stone
- Resource Type
- Journal article
- Publication Details
- Proceedings of the National Academy of Sciences - PNAS, Vol.108(34), pp.E569-E576
- DOI
- 10.1073/pnas.1108918108
- PMID
- 21825139
- PMCID
- PMC3161526
- NLM abbreviation
- Proc Natl Acad Sci U S A
- ISSN
- 0027-8424
- eISSN
- 1091-6490
- Publisher
- United States
- Grant note
- R01 EY016822 / NEI NIH HHS DP2 OD007483 / NIH HHS EY017451 / NEI NIH HHS R01 EY016822-06 / NEI NIH HHS R01 EY017451 / NEI NIH HHS EY016822 / NEI NIH HHS R01 EY016822-07 / NEI NIH HHS DP2 OD007483-01 / NIH HHS R01 EY016822-08 / NEI NIH HHS
- Language
- English
- Date published
- 08/23/2011
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Electrical and Computer Engineering; Stead Family Department of Pediatrics; Iowa Neuroscience Institute; Medical Genetics and Genomics; Center for Bioinformatics and Computational Biology; Ophthalmology and Visual Sciences
- Record Identifier
- 9983980069002771
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