A chemical biology toolbox for RNA post-transcriptional modification and capture

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Abstract RNA serves as the key intermediate for carrying genetic information and translating functional proteins, and numerous types of non-coding RNAs have also been recently discovered that play critical roles in cellular processes. While RNAs are directly transcribed from DNA, their sequence, function, and translation efficiency are all actively modulated in the cell though post-transcriptional localization and editing. These processes are essential for development and cellular function and are dysregulated in many diseases. Despite the importance of post-transcriptional localization and modification, significant gaps remain in our understanding of the timing, mechanisms, and regulation of these events. Several methods have been reported that enable researchers to label and image specific RNAs in living cells or pull-down and sequence edited transcripts from cell lysates. However, significant technological limitations still exist, and addressing these gaps holds promise for the development of new therapeutics and diagnostics. We have previously developed and implemented new methods for covalent labeling and imaging of specific RNAs in cells as well as pull-down and enrichment of A- to-I edited transcripts from cellular RNA. The future directions for our program include expanding this chemical biology toolbox to provide researchers with new and improved technologies for probing these important cellular processes. Additionally, recent studies have offered groundbreaking evidence for the hypothesis that asymmetric localization of RNA serves as a mechanism for regulating editing. Thus, a significant focus of the proposed research is to combine our imaging and editing-specific pull-down methods to advance understanding of the interplay between these two processes. Together, this research aims to serve the scientific community by providing insight into the regulation of RNA localization and modification as well as developing and disseminating robust and well-validated technologies for studying and modulating biological systems.