Niu Yuping, Huang Sexin, Xu Peiwen, Li Jie, Gao Ming, Chen Xiaowei, Chu Hongxia, Gao Yuan
Center for Reproductive Medicine Research, Shandong University; National Research Center for Assisted Reproductive Technology and Reproductive Genetics; Key Laboratory for Reproductive Endocrinology of Ministry of Education, Jinan, Shandong 250001, China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2021 Jun 10;38(6):553-556. doi: 10.3760/cma.j.cn511374-20200211-00072.
To explore the genetic basis for a patient with tuberous sclerosis complex.
Genomic DNA was extracted from peripheral blood samples from members of his family and 100 unrelated healthy controls. The proband was subjected to next-generation sequencing, and candidate variant was confirmed by multiple ligation-dependent probe amplification (MLPA) and Sanger sequencing. Reverse transcription-PCR (RT-PCR) was carried out to determine the relative mRNA expression in the proband.
The patient was found to harbor a c.2355+1G>C splicing variant of the TSC2 gene. Sequencing of cDNA confirmed that 62 bases have been inserted into the 3' end of exon 21, which has caused a frameshift producing a truncated protein.
The novel splicing variant c.2355+1G>C of the TSC2 gene probably underlay the TSC in the proband. Above finding has expanded the variant spectrum of TSC2 and provided a basis for preimplantation genetic testing and/or prenatal diagnosis.
探究一名结节性硬化症患者的遗传基础。
从其家族成员的外周血样本以及100名无关健康对照者中提取基因组DNA。对先证者进行二代测序，并通过多重连接依赖探针扩增（MLPA）和桑格测序确认候选变异。进行逆转录聚合酶链反应（RT-PCR）以确定先证者中相对mRNA的表达。
发现该患者携带TSC2基因的c.2355+1G>C剪接变异。cDNA测序证实62个碱基已插入外显子21的3'末端，这导致了移码，产生了截短的蛋白质。
TSC2基因新的剪接变异c.2355+1G>C可能是先证者结节性硬化症的病因。上述发现扩展了TSC2的变异谱，为植入前基因检测和/或产前诊断提供了依据。
