transgenic expression of UPRT as a novel tool for tagging RNA in specific tissues of the mouse

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Summary (Lay)RNA is synthesised using information encoded by DNA and it mediates many of the functions of the genome as well as producing proteins. Understanding the regulation of RNA is a priority for the understanding of cell function. Like DNA sequences, RNA sequences are made up for four bases, but, unlike DNA, RNA contains the nucleoside uridine which is composed of the pyrimidine uracil. Uracil is not incorporated into the RNA of mouse or human cells as the required enzymes for this pathway are not encoded in the mammalian genome. Consequently, controlled expression of the enzyme uracil phosphoribosyltransferase (UPRT) which converts uracil into uridine monophosphate allows the incorporation of uracil in cell-type specific or temporally controlled manner labelling of RNA. We have generated a transgenic mouse that expresses UPRT in a controllable way and we want to demonstrate the broad utility of this as a tool for purification of RNA or the identification of RNA protein interactions in rare cell subsets in complex mixtures of cells. This is an improvement because it will open access to the RNA makeup of some cell types that cannot be easily purified. Furthermore, the act of purification is known to change the quantity and quality of the RNA within the cell. Thus by reducing the numbers of manipulations a more accurate readout of RNA content or RNA-protein interactions will be obtained.