ResearchIn-Press PreviewGeneticsOphthalmology Open Access | 10.1172/jci.insight.183902
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Nakamichi, K. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Huey, J. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Sangermano, R. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Place, E. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Bujakowska, K. in: JCI | PubMed | Google Scholar |
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Marra, M. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Everett, L. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Yang, P. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Chao, J. in: JCI | PubMed | Google Scholar
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Van Gelder, R. in: JCI | PubMed | Google Scholar |
1Department of Ophthalmology, University of Washington, Seattle, United States of America
2Department of Ophthalmology, Harvard Medical School, Boston, United States of America
3Casey Eye Institute, Oregon Health & Science University, Portland, United States of America
Find articles by Mustafi, D. in: JCI | PubMed | Google Scholar
Published September 12, 2024 - More info
Despite advances in sequencing technologies, a molecular diagnosis remains elusive in many Mendelian disease patients. Current short-read clinical sequencing approaches cannot provide chromosomal phase information or epigenetic information without further sample processing, which is not routinely done and can result in an incomplete molecular diagnosis in patients. The ability to provide phased genetic and epigenetic information from a single sequencing run would improve the diagnostic rate of Mendelian conditions. Here we describe Targeted Long-read Sequencing of Mendelian Disease genes (TaLon-SeqMD) using a real-time adaptive sequencing approach. Optimization of bioinformatic targeting enabled selective enrichment of multiple disease-causing regions of the human genome. Haplotype-resolved variant calling and simultaneous resolution of epigenetic base modification could be achieved in a single sequencing run. The TaLon-SeqMD approach was validated in a cohort of 18 subjects with previous genetic testing targeting 373 inherited retinal disease (IRD) genes, yielding the complete molecular diagnosis in each case. This approach was then applied in two IRD cases with inconclusive testing, which uncovered non-coding and structural variants that were difficult to characterize by standard short-read sequencing. Overall, these results demonstrate TaLon-SeqMD as an approach to provide rapid phased-variant calling to provide the molecular basis of Mendelian diseases.