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Nucleic Acids Research发表线粒体测序技术

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摘要 : 线粒体是人类细胞将糖转化成能量的重要工厂。西奈山伊坎医学院的科学家们开发了一种新测序技术,可以帮助人们理解线粒体基因对个体患病风险的影响,比如糖尿病、心脏病和癌症。这项研究发表在近期的Nucleic Acids Research杂志上。

Nucleic Acids Research发表线粒体测序技术

线粒体是人类细胞将糖转化成能量的重要工厂。西奈山伊坎医学院的科学家们开发了一种新测序技术,可以帮助人们理解线粒体基因对个体患病风险的影响,比如糖尿病、心脏病和癌症。这项研究发表在近期的Nucleic Acids Research杂志上。

在年龄、体重和坏习惯(比如吸烟)相同的人中,有些会生病有些却不会。研究者们一直对这一点感到迷惑,并尝试通过饮食和核基因来确定个体的患病风险。但这样的分析遗漏了线粒体基因(mtDNA)的差异,mtDNA遗传自我们的母亲,每个细胞都有这样的DNA。

近年来研究表明,线粒体发生错误会引起严重的疾病。然而由于mtDNA的异质性很大,分析mtDNA变异对个体疾病风险的影响非常复杂。

这项研究展示了首个可以准确鉴定mtDNA异质性的测序技术,Mseek。细胞里的mtDNA含量很少,过去这方面一直没有什么进展。人们意识到了mtDNA的异质性,但却无法再单细胞水平上建立决定性的理论。

“研究者们一直在寻找能够准确测序mtDNA的低成本方法,”西奈山伊坎医学院的助理教授Ravi Sachidanandam博士说。“现在我们做到了这一点。Mseek可以鉴定线粒体中的功能紊乱,使mtDNA成为了有用的生物学指标。此外,人们还可以将mtDNA作为潜在的靶标,治疗癌症和多种遗传疾病。”

据Dr. Sachidanandam介绍,新测序技术达到了空前的灵敏性、特异性和测序深度,能够得到高纯度的mtDNA,检测不同版本的mtDNA或mtDNA变体。

Mseek主要是利用酶清除细胞核DNA,只留下线粒体DNA进行测序。研究人员将Mseek用于多个细胞系,鉴定了不同细胞样本中的mtDNA“指纹”。他们还通过追踪mtDNA变体发现,mtDNA异质性在单细胞水平上是稳定维持的。

“我们曾经推测,异质性可以通过细胞间的mtDNA交换稳定下来,”Sachidanandam说。“结果显示,mtDNA交换的确可行,异质性能够通过这一机制维持稳定。这一技术为人们提供了一个新平台,可以研究正常和疾病状态下mtDNA异质性特征。未来也许可以利用交换机制进行治疗,将坏mtDNA换成好mtDNA。”

原文链接:Stable heteroplasmy at the single-cell level is facilitated by intercellular exchange of mtDNA

Eukaryotic cells carry two genomes, nuclear (nDNA) and mitochondrial (mtDNA), which are ostensibly decoupled in their replication, segregation and inheritance. It is increasingly appreciated thatheteroplasmy, the occurrence of multiple mtDNA haplotypes in a cell, plays an important biological role, but its features are not well understood. Accurately determining the diversity of mtDNA has been difficult, due to the relatively small amount of mtDNA in each cell (<1%of the total DNA), the intercellular variability of mtDNA content andmtDNA pseudogenes (Numts) in nDNA. To understand the nature ofheteroplasmy, we developed Mseek, a novel technique to purify and sequence mtDNA. Mseek yields high purity (>90%) mtDNA and its ability to detect rare variants is limited only by sequencing depth, providing unprecedented sensitivity and specificity. Using Mseek, we confirmed theubiquity of heteroplasmy by analyzing mtDNA from a diverse set of cell lines and human samples. Applying Mseek to colonies derived from single cells, we find heteroplasmy is stably maintained in individual daughter cells over multiple cell divisions. We hypothesized that the stability ofheteroplasmy could be facilitated by intercellular exchange ofmtDNA. We explicitly demonstrate this exchange by co-culturing cell lines with distinct mtDNA haplotypes. Our results shed new light on themaintenance of heteroplasmy and provide a novel platform to investigate features of heteroplasmy in normal and diseased states.

作者:Anitha D. Jayaprakas 点击:

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