open access

https://hdl.handle.net/2115/90104

Cas9-guided haplotyping of three truncation variants in autosomal recessive disease

英語

nanopore sequencing

2022-07

John Wiley & Sons

Human mutation

43(7)

877-881

An autosomal recessive disease is caused by biallelic loss-of-function mutations. However, when more than two disease-causing variants are found in a patient's gene, it is challenging to determine which two of the variants are responsible for the disease phenotype. Here, to decipher the pathogenic variants by precise haplotyping, we applied nanopore Cas9-targeted sequencing (nCATS) to three truncation COL7A1 variants detected in a patient with recessive dystrophic epidermolysis bullosa (EB). The distance between the most 5 ' and 3 ' variants was approximately 19 kb at the level of genomic DNA. nCATS successfully demonstrated that the most 5 ' and 3 ' variants were located in one allele while the variant in between was located in the other allele. Interestingly, the proband's mother, who was phenotypically intact, was heterozygous for the allele that harbored the two truncation variants, which could otherwise be misinterpreted as those of typical recessive dystrophic EB. Our study highlights the usefulness of nCATS as a tool to determine haplotypes of complicated genetic cases. Haplotyping of multiple variants in a gene can determine which variant should be therapeutically targeted when nucleotide-specific gene therapy is applied.

学術雑誌論文

Accepted Manuscript

This is the peer reviewed version of the following article: Natsuga, K., Furuta, Y., Takashima, S., Nohara, T., Huang, H.-Y., Shinkuma, S., Nakamura, H., Katsuda, Y., Higashi, H., Hsu, C.-K., Fukushima, S., & Ujiie, H. (2022). Cas9-guided haplotyping of three truncation variants in autosomal recessive disease. Human Mutation, 43, 877– 881, which has been published in final form at 10.1002/humu.24385. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. This article may not be enhanced, enriched or otherwise transformed into a derivative work, without express permission from Wiley or by statutory rights under applicable legislation. Copyright notices must not be removed, obscured or modified. The article must be linked to Wiley’s version of record on Wiley Online Library and any embedding, framing or otherwise making available the article or pages thereof by third parties from platforms, services and websites other than Wiley Online Library must be prohibited.

1059-7794