講演情報
[O29-3]iGONAD法を用いた迅速な疾患モデルマウス作出による遺伝性疾患に見出された病的意義不明なゲノムバリアント (VUS)の機能解明
○林 深, 鈴木 康予, 加藤 君子, 福士 大輔, 山田 憲一郎 (愛知県医療療育総合センター 発達障害研究所 遺伝子医療研究部)
The dramatic development of genetic analysis in the last two decades, e.g. next-generation sequencing or microarray analysis, has identified a number of causative genes and pathogenic variants in genetic disorders. However, such high-throughput genetic analyses have also found enormous variants of uncertain significance (VUS) concurrently and it remains unsolved problems how to interpret etiological significance of VUS. Although in vitro analysis like functional analysis of mutant protein or in silico estimation may provide evidence for understanding VUS, they are sometimes inadequate to clarify a correlation between VUS and phenotype. To resolve the problem, we have attempted to generate rapidly model mice mimicking the targeting VUS precisely using the iGONAD method. The method generates transgenic mice ate least in 19 days by intraoviductal inducing of genome-editing reagents into fertilized ovum directly. To date, we have generated ten strains of intended transgenic mice of ten variants in five genes. For example, recently we found a familial epilepsy with a variant at a splice site of TENM4, while missense variants of TENM4 are known to cause an essential tremor. In order to explain their etiology, we generated mice reproducing the mutation at the splice site. Transcript and protein analyses of the mice brains showed a short form of Tenm4 protein with exon skipping, which may change a form of TENM4 protein in a neural cell. Now we are assessing phenotype and convulsant property of the mice. Our current methodology is useful in understanding VUS in genetic disorders.