Combining viral and ribosomal mRNA capture technologies to develop a versatile system for neuronal transcriptome profiling

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It is very difficult to study the function(s) of specific groups of neurons because the brain contains thousands of different cell types that are juxtaposed and interconnected and vary according to their size, shape, projections and function. Recently a method has been developed that allows all the molecules (messenger RNA) that are being translated into proteins to be in defined in specific neuronal populations. This is a very powerful tool as for the first time the sets of genes that are governing neuronal function (e.g. those controlling the formation of memories) and those that are altered with age and in human neurodegenerative and neuropsychiatric illnesses can be identified. However, this method is dependent on costly transgenic mice lines, and each time a new scientific question is asked a new mouse line needs to be generated. The technique itself is quite complex, and time consuming due to the need to develop and breed (and in many cases cross breed) one or more mouse lines. These considerations severely limit the availability of this technique to researchers and instead a combination of less powerful approaches must be used. In this study we are combining the expertise of Takeda and the University of Bristol to develop two new methods. These methods use viral vectors instead of transgenic mice (called viral TRAP) and allow neurons to be profiled with a speed, precision and versatility not previously possible with transgenic mice alone. These viral TRAP methods will also result in fewer animals being used by researchers. The viral TRAP technique will be used by the University of Bristol to identify the genes and proteins a family of RNA binding proteins called scaffold attachment factors (SAF) regulate. In particular this information will be used to understand how SAF proteins govern the processes that regulate memory formation and ageing. The viral TRAP technology will enable Takeda to perform detailed profiling experiments without the need for lengthy, expensive and animal-intensive transgenic programs. Importantly as the viral TRAP technique is portable to other models that Takeda currently use the technology will provide a novel platform that will greatly facilitate both basic biology and future pharmacological response studies. Ultimately, the objective of Takeda is to use this technology to further the understanding of neuronal gene regulation and homeostasis in response to challenge, and to use these insights to identify novel targets for central nervous system disorders.