Cas9‐guided haplotyping of three truncation variants in autosomal recessive disease
Cas9‐guided haplotyping of three truncation variants in autosomal recessive disease
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Cas9引导的常染色体隐性遗传病三种截断变异的单倍型分析
DOI：
10.1002/humu.24385
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发表时间：
2022
期刊：
影响因子：
3.9
通讯作者：
Ujiie Hideyuki
中科院分区：
文献类型：
--
作者：
Natsuga Ken;Furuta Yoshikazu;Takashima Shota;Nohara Takuma;Huang Hsin‐Yu;Shinkuma Satoru;Nakamura Hideki;Katsuda Yousuke;Higashi Hideaki;Hsu Chao‐Kai;Fukushima Satoshi;Ujiie Hideyuki
常染色体隐性遗传病是由双等位基因功能丧失突变引起的。然而，当在患者的基因中发现两种以上的致病变异时，确定哪两种变异导致疾病表型是具有挑战性的。在这里，为了通过精确的单体型分析来解读致病性变体，我们将纳米孔Cas9靶向测序（nCATS）应用于在隐性营养不良性大疱性表皮病（EB）患者中检测到的三种截短COL7A1变体。在基因组DNA水平上，大多数5′和3′变异体之间的距离约为19kb。nCATS成功地证明了大多数5′和3′变异位于一个等位基因中，而介于两者之间的变异位于另一个等位基因中。有趣的是，先证者的母亲，谁是表型完整，是杂合子的等位基因，窝藏两个截断变异，这可能会被误解为典型的隐性营养不良EB。我们的研究强调了nCATS作为确定复杂遗传病例单倍型的工具的有用性。基因中的多个变体的单体型分析可以确定当应用核苷酸特异性基因治疗时哪个变体应该是治疗靶向的。
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 truncationCOL7A1variants 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.
DOI：
10.1073/pnas.92.15.6971
发表时间：
1995-07-18
影响因子：
11.1
作者：
NEGRONI, M;RICCHETTI, M;BUC, H
通讯作者：
BUC, H
通过 Cas9 靶向长读长测序检测大疱性表皮松解症中的回复嵌合体
影响因子：
3.9
作者：
Natsuga Ken;Furuta Yoshikazu;Takashima Shota;Nohara Takuma;Kosumi Hideyuki;Mai Yosuke;Higashi Hideaki;Ujiie Hideyuki
通讯作者：
Ujiie Hideyuki