1. Long read mitochondrial genome sequencing using Cas9-guided adaptor ligation.
- Author
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Vandiver AR, Pielstick B, Gilpatrick T, Hoang AN, Vernon HJ, Wanagat J, and Timp W
- Subjects
- Base Sequence, CRISPR-Cas Systems, DNA, Mitochondrial genetics, High-Throughput Nucleotide Sequencing methods, Humans, Mitochondria genetics, Sequence Analysis, DNA methods, Genome, Mitochondrial
- Abstract
The mitochondrial genome (mtDNA) is an important source of disease-causing genetic variability, but existing sequencing methods limit understanding, precluding phased measurement of mutations and clear detection of large sporadic deletions. We adapted a method for amplification-free sequence enrichment using Cas9 cleavage to obtain full length nanopore reads of mtDNA. We then utilized the long reads to phase mutations in a patient with an mtDNA-linked syndrome and demonstrated that this method can map age-induced mtDNA deletions. We believe this method will offer deeper insight into our understanding of mtDNA variation., (Copyright © 2022 Elsevier B.V. and Mitochondria Research Society. All rights reserved.)
- Published
- 2022
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