Institute of Forensic Medicine, Medical Center, University of Freiburg and Faculty of Medicine, University of Freiburg, Freiburg 79104, Germany.
Institute of Legal Medicine, Medical University of Innsbruck, Innsbruck 6020, Austria.
Nucleic Acids Res. 2021 Feb 22;49(3):1517-1531. doi: 10.1093/nar/gkaa1271.
The maternal mode of mitochondrial DNA (mtDNA) inheritance is central to human genetics. Recently, evidence for bi-parental inheritance of mtDNA was claimed for individuals of three pedigrees that suffered mitochondrial disorders. We sequenced mtDNA using both direct Sanger and Massively Parallel Sequencing in several tissues of eleven maternally related and other affiliated healthy individuals of a family pedigree and observed mixed mitotypes in eight individuals. Cells without nuclear DNA, i.e. thrombocytes and hair shafts, only showed the mitotype of haplogroup (hg) V. Skin biopsies were prepared to generate ρ° cells void of mtDNA, sequencing of which resulted in a hg U4c1 mitotype. The position of the Mega-NUMT sequence was determined by fluorescence in situ hybridization and two different quantitative PCR assays were used to determine the number of contributing mtDNA copies. Thus, evidence for the presence of repetitive, full mitogenome Mega-NUMTs matching haplogroup U4c1 in various tissues of eight maternally related individuals was provided. Multi-copy Mega-NUMTs mimic mixtures of mtDNA that cannot be experimentally avoided and thus may appear in diverse fields of mtDNA research and diagnostics. We demonstrate that hair shaft mtDNA sequencing provides a simple but reliable approach to exclude NUMTs as source of misleading results.
线粒体 DNA（mtDNA）的母系遗传模式是人类遗传学的核心。最近，有证据表明，在三个患有线粒体疾病的家系个体中，mtDNA 存在双亲遗传。我们对一个家系的 11 名具有血缘关系的和其他相关的健康个体的多个组织进行了直接 Sanger 和大规模平行测序，在 8 名个体中观察到混合的单体型。没有核 DNA 的细胞，即血小板和头发，仅显示单倍型（hg）V 的单体型。我们准备皮肤活检以生成缺乏 mtDNA 的 ρ°细胞，对其进行测序得到 hg U4c1 单体型。通过荧光原位杂交确定 Mega-NUMT 序列的位置，并使用两种不同的定量 PCR 检测来确定贡献的 mtDNA 拷贝数。因此，在 8 名具有血缘关系的个体的多个组织中存在重复的、完整的与单倍型 U4c1 匹配的线粒体 Mega-NUMTs 的证据。多拷贝的 Mega-NUMTs 模拟了无法通过实验避免的 mtDNA 混合物，因此可能出现在 mtDNA 研究和诊断的各个领域。我们证明，头发轴 mtDNA 测序提供了一种简单但可靠的方法来排除 NUMTs 作为产生误导结果的来源。
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