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

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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.