Role of mrna internal 2′-o methylation in post-transcriptional regulation - title: role of mrna internal 2′-o methylation in post-transcriptional regulation project summary/abstract: emerging evidence…

The National Institute of General Medical Sciences awarded Northwestern University $440,000 on April 3, 2026, under the Biomedical Research and Research Training program (CFDA 93.859) to investigate the role of mRNA internal 2-O methylation in post-transcriptional regulation.

The research combines nanopore direct RNA sequencing and machine learning methods to characterize how 2-O methylation (NM) governs mRNA function and metabolism. Three complementary projects are proposed: the first identifies HnRNPF/H1 as downstream effector proteins that mediate how NM modification remodels mRNA splicing outcomes globally; the second uncovers the mechanism underlying crosstalk between NM and N6-methyladenosine (M6A) modification and characterizes the impact of dual methylation on mRNA fate; and the third addresses dysregulation of NM in disease contexts. The research employs comprehensive proteomic, molecular, and multi-omics approaches.

Northwestern University's Sponsored Research Division is the recipient. Work is performed in Chicago, Illinois. The award period runs through December 31, 2030. This is a Project Grant, the standard assistance type under NIGMS's portfolio-based funding approach emphasizing investigator-initiated biomedical research that advances understanding of biological processes underlying disease.

Role of mrna internal 2′-o methylation in post-transcriptional regulation - title: role of mrna internal 2′-o methylation in post-transcriptional regulation project summary/abstract: emerging evidence has proved the pivotal roles of RNA modifications in the control of various fundamental bioprocesses as well as diseases. As a prevalent modification type in most RNA species, 2′-o methylation (NM) could present at all four ribonucleosides to affect RNA structure, stability, and interactions. In sharp contrast to the extensively studied rrna NM and 5′-cap mrna NM, NM occurred at mrna internal sites were only reported until recently, while its biological functions and underlying mechanisms remain underexplored. By combining nanopore direct RNA-SEQ (nanopore-SEQ) with our newly developed machine learning method, we recently identified thousands of novel mrna NM sites in human cells at single-base resolution and characterized fibrillarin (FBL) as a major 2′-o methyltransferase (mtase) to install this modification. Despite these advances, little is known about the regulatory network by which NM governs mrna functions and metabolism, and how NM is manipulated to ensure the precise and synchronous regulation of gene expression. In our current studies, we propose three independent yet complementary projects to resolve this knowledge GAP. By utilizing comprehensive proteomic, molecular, and multi-omics approaches, the first project will identify hnrnpf/H1 as distinct NM downstream effector proteins and elucidate how this NM & effector signaling remodels the splicing outcome of NM-modified mrnas globally. In addition, our nanopore-SEQ results have revealed the evident CO-occurrence of NM with N6-methyladenosine (M6A), the most abundant RNA modifications in mammalian cells. Therefore, our second project will uncover the mechanism underlying the crosstalk between NM and M6A modification and elucidate the impact of NM & M6A dual methylation on mrna fate. Dysregulation of NM is frequently linked to multiple neurodevelopmental disorders. Hence, our last project will dedicate to discover the functional relevance of mrna NM on pluripotency maintenance and neuronal fate decision. Overall, our studies will gain novel insights into the biological functions of mrna internal NM and pioneer the NM-targeting technological innovations to benefit human health.