Nanobaths for DNA analysis

用于 DNA 分析的纳米浴

• 批准号: BB/M018962/1
• 负责人: Tracy Melvin
• 金额: $ 17.95万
• 依托单位: University of Southampton
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM018962%2F1
• 关键词: Nanobaths; DNA; analysis

The human genome contains 3164.7 million nucleic acid bases (adenine, guanine, cytosine, thymine) and it is estimated that the length of all the DNA strands in a single cell (if all the strands were placed end to end) is about two metres. The human genome sequence was completed at the start of the millennium; this resulted in significant public and scientific interest in understanding the DNA sequence 'code' and how it is 'translated'. The sequence of the genome provides information about our ancestry, hereditary diseases, our features (such as eye, skin or hair colour) and our physiological 'make-up'. Despite the fact that the human genome was sequenced a decade ago and better DNA sequencing methods have been since developed, simpler, cheaper, faster DNA sequence analysis methods which do not necessarily provide the full genome sequence but detect specific differences in the sequence is an important goal. Thus to obtain information that is required without evaluating the whole sequence - essentially just like checking the key quotes from a Shakespeare play without reading the whole book 'cover to cover'.In our view, what is needed is a small scale technology, something that works like a thermal reader, where a tiny read head is scanned past the stored information (the DNA strand) and the information (the differences in the sequence) is read directly without need for any complex processing of the genomic DNA molecule. We propose to flow DNA molecules through a nanocapillary with a light activated heater that will act as a 'read head' and detect and identify variations in the DNA sequence. The DNA is confined in channels meaning that the long DNA strand which can be microns long does not travel through all 'noted up'. Identifying variations in the DNA base sequence in this way will be very simple and fast, and we believe capable of detecting modifications to particular bases - notably sequence variations inherited from parents, from damage leading to cancers, from environmental or cellular processes which can control the switching on and off of genes. These sorts of techniques are crucial to obtain a better understand of the genetics of all organisms, not just humans, and a fast, cheap DNA analysis method will be able to answer many more questions as well as also provide a simple fast diagnostic tool. These studies will provide proof of concept data appropriate to demonstrate the potential of nanocapillaries for the first time for integration with optical approaches for single molecule interrogation in confined spaces. We believe these capillaries could be developed to provide simpler easier tools for other diagnostic applications.

人类基因组包含31.647亿个核酸碱基（腺嘌呤、鸟嘌呤、胞嘧啶、胸腺嘧啶），据估计，单个细胞中所有DNA链的长度（如果所有链首尾相连）约为2米。人类基因组序列在千禧年开始时完成，这导致了公众和科学界对理解DNA序列“代码”以及如何“复制”的重大兴趣。基因组序列提供了关于我们的祖先、遗传性疾病、我们的特征（如眼睛、皮肤或头发的颜色）和我们的生理“组成”的信息。尽管人类基因组在十年前就被测序，并且此后已经开发出更好的DNA测序方法，但更简单、更便宜、更快的DNA序列分析方法（不一定提供完整的基因组序列，但检测序列中的特定差异）是一个重要目标。因此，在不评估整个序列的情况下获得所需的信息-本质上就像检查莎士比亚戏剧中的关键引文而不“从头到尾”阅读整本书一样。在我们看来，需要的是一种小规模的技术，类似于热阅读器的东西，一个微小的读取头扫描存储的信息直接读取基因组DNA（DNA链）和信息（序列中的差异），而不需要对基因组DNA分子进行任何复杂的处理。我们建议让DNA分子流过具有光激活加热器的纳米毛细管，该加热器将充当“读取头”并检测和识别DNA序列中的变化。DNA被限制在通道中，这意味着可以是微米长的长DNA链不会穿过所有的“记录”。以这种方式识别DNA碱基序列中的变异将非常简单和快速，我们相信能够检测特定碱基的修饰-特别是从父母继承的序列变异，导致癌症的损伤，可以控制基因开关的环境或细胞过程。这些技术对于更好地了解所有生物体的遗传学至关重要，而不仅仅是人类，快速，廉价的DNA分析方法将能够回答更多的问题，并提供简单快速的诊断工具。这些研究将提供适当的概念数据证明，以证明纳米胶囊的潜力，第一次与光学方法集成，用于在有限空间中进行单分子询问。我们相信这些毛细管可以被开发为其他诊断应用提供更简单更容易的工具。