Multiplex Bioorthogonal Labelling of Nucleic Acids: A Tool for Super-Resolution Imaging

了解哺乳动物细胞在正常发育和疾病中的功能是生命科学研究的核心，也是发现新药物和治疗方法的必要条件。许多生物系统非常复杂，包含许多不同的组成部分。为了剖析这个谜题的各个部分，有必要使用可用于标记各个组件的特定标签。在过去的30年里，许多不同的方法被用来标记拼图的特定部分，通常是用不同的颜色。细胞中的一些关键因素是由蛋白质组成的单个组件，类似于汽车的部件。这些不同的蛋白质可以用称为抗体的特定试剂进行标记，这些抗体可以用不同的颜色进行标记，并通过显微镜进行可视化，以探索它们在3D空间中如何相互关联。类似地，这种方法可以在分子水平上使用，以询问这些组分是否相互作用。有非常好的工具来分析蛋白质，相比之下，在细胞中标记DNA和RNA要困难得多。DNA编码我们的遗传信息，并与蛋白质仔细折叠，以保护它免受损害，并调节信息的访问方式。为了制造蛋白质，细胞将储存在DNA中的信息复制到称为RNA的临时代码中。反过来，RNA被用作制造蛋白质的配方。RNA和DNA一起被描述为核酸，尽管它们是关键成分，但很难单独标记它们，因为它们看起来非常相似。我们最近发现，在细胞中，DNA与RNA和蛋白质一起包装成一个类似于渔网的网，这对调节细胞的功能非常重要，但我们意识到我们缺乏研究这种关系的工具。了解这些相互作用至关重要，因为制造网状物的蛋白质之一通常在癌症中发生突变，而这种蛋白质的基因突变会导致儿童的发育缺陷。因此，为了了解蛋白质、DNA和RNA如何协同工作，我们将开发一种新的试剂调色板，使我们能够同时用不同的颜色标记细胞中的这些不同成分。这种多重方法将适用于许多不同的研究，但将至关重要地使人们能够了解单个成分如何共同调节DNA，以及在疾病中如何出错。要剖析生命的复杂运作，必须使用创新的方法。为了实现我们的目标，我们组建了一个由化学家和分子生物学家组成的团队，他们可以一起制造新的工具来了解生命是如何运作的。

Understanding how mammalian cells function in normal development and disease is central to life science research, and is necessary to discover new drugs and treatments. Many biological systems are extremely complex and comprise many different components. To dissect the individual parts of this puzzle it is necessary to use specific labels that can be used to mark individual components. Over the last 30-years many different approaches have been used for labelling specific parts of the puzzle, often in different colours. Some of the key factors in a cell are the individual components made of proteins and analogous to the parts of a car. These different proteins can be labelled with specific reagents called antibodies that can be tagged in different colours and visualised by microscopy to explore how they relate to each other in 3D space. Similarly this approach can be used on a molecular level to ask if these components interact with each other. There are very good tools for analysing proteins, in contrast it is much more difficult to label DNA and RNA in cells. DNA encodes our genetic information and is carefully folded with proteins to protect it from damage and to regulate how the information is accessed. To make proteins cells copy the information stored in DNA into a temporary code called RNA. In turn RNA is used as a recipe to make proteins. Together RNA and DNA are described as nucleic acids and although these are critical components it is difficult to separately label them, as they look very similar. We have recently shown that in cells, DNA is packaged with RNA and proteins to make a mesh, similar to a fishing net, that is important for regulating how cells function, but we realised that we lacked the tools to investigate this relationship. Understanding these interactions is crucial as one of the proteins for making the mesh is often mutated in cancer, and genetic mutations in this protein cause developmental defects in children. Therefore, to understand how proteins, DNA and RNA work together we will develop a new palette of reagents that will allow us to mark these different components in cells in different colours simultaneously. This multiplex approach will be applicable to many different studies but will critically enable use to understand how individual components work together in regulating DNA and how in disease this can go wrong. To dissect the complex workings of life it is necessary to use innovative methods. To achieve our goals we have assembled a team of chemists and molecular biologists who together can make new tools for understanding how life works.