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Detection of revertant mosaicism in epidermolysis bullosa through Cas9-targeted long-read sequencing
| Title: | Detection of revertant mosaicism in epidermolysis bullosa through Cas9-targeted long-read sequencing |
| Authors: | Natsuga, Ken Browse this author →KAKEN DB | | Furuta, Yoshikazu Browse this author | | Takashima, Shota Browse this author | | Nohara, Takuma Browse this author | | Kosumi, Hideyuki Browse this author | | Mai, Yosuke Browse this author | | Higashi, Hideaki Browse this author | | Ujiie, Hideyuki Browse this author →KAKEN DB |
| Keywords: | epidermolysis bullosa | | intragenic crossover | | long-read sequencing | | revertant mosaicism |
| Issue Date: | 1-Apr-2022 |
| Publisher: | John Wiley & Sons |
| Journal Title: | Human mutation |
| Volume: | 43 |
| Issue: | 4 |
| Start Page: | 529 |
| End Page: | 536 |
| Publisher DOI: | 10.1002/humu.24331 |
| Abstract: | Revertant mosaicism (RM) is a phenomenon in which inherited mutations are spontaneously corrected in somatic cells. RM occurs in some congenital skin diseases, but genetic validation of RM in clinically revertant skin has been challenging, especially when homologous recombination (HR) is responsible for RM. Here, we introduce nanopore Cas9-targeted sequencing (nCATS) for identifying HR in clinically revertant skin. We took advantage of compound heterozygous COL7A1 mutations in a patient with recessive dystrophic epidermolysis bullosa who showed revertant skin spots. Cas9-mediated enrichment of genomic DNA (gDNA) covering the two mutation sites (>8 kb) in COL7A1 and subsequent MinION sequencing successfully detected intragenic crossover in the epidermis of the clinically revertant skin. This method enables the discernment of haplotypes of up to a few tens of kilobases of gDNA. Moreover, it is devoid of polymerase chain reaction amplification, which can technically induce recombination. We, therefore, propose that nCATS is a powerful tool for understanding complicated gene modifications, including RM. |
| Rights: | This is the peer reviewed version of the following article: Natsuga, K., Furuta, Y., Takashima, S., Nohara, T., Kosumi, H., Mai, Y., Higashi, H., & Ujiie, H. (2022). Detection of revertant mosaicism in epidermolysis bullosa through Cas9-targeted long-read sequencing. Human Mutation, 43, 529– 536 which has been published in final form at doi:10.1002/humu.24331. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/88789 |
| Appears in Collections: | 医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 夏賀 健
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