Elucidating the biological role of the 'epitranscriptome'
Školitel: Dr. Mary O'Connell
Instituce: Masaryk University, Faculty of Science
O mém projektu
By combining one genetic model system (mouse models) with biochemical and cell biological experiments and nucleotide analysis by liquid chromatography MS (LC-MS) my study will take a comprehensive look at the occurrence and interplay of different types of RNA modifications. This will be the first time that multiple modifications will be considered simultaneously. Recently there has been a surge of interest in RNA modification; the ‘epitranscriptome’, has been established as a major new field. The extent, diversity and consequence of RNA modifications in the transcriptome are unknown. The modifications studied so far can influence translation, folding, binding of proteins and degradation of RNA. The diseases that they have been implicated in range from cancer to neurodegeneration and even obesity. It is impossible to easily discern RNA modifications as RNA Seq use reverse transcriptase that fails to distinguish between canonical and modified bases. Consequently, most researchers focus on immune-precipitations or chemical reactions targeting just one type of modification. In contrast, we use liquid chromatography-mass spectrometry (LC-MS), the gold standard to detect base modifications. This proposal will focus on inosine and the ADAR enzymes as inosine is one of the most abundant RNA modifications in the transcriptome. In addition we propose to examine the interplay between inosine and some of the other more abundant modifications and determine how they can influence each other.Cell culture will be used to examine various parameters such as stress, induction of the immune response and cell aging. The goal will be to elucidate if there is an orchestrated response in RNA modification to a changing environment.