# A universal approach for determining three-dimensional RNA structures

> **NIH NIH R21** · NEW YORK UNIVERSITY SCHOOL OF MEDICINE · 2024 · $169,500

## Abstract

Summary
RNA molecules participate in the most fundamental cellular processes implicated in human disease. Many
RNAs contain structured modules that make critical contributions to RNA functions and represent attractive
drug targets, especially for diseases without a cure and associated with “undruggable” proteins. Knowledge of
the three-dimensional structures of these RNAs would help to understand the mechanism of the RNA function
and could greatly accelerate drug discovery efforts. However, traditional methods for RNA structure
determination, X-ray crystallography and NMR spectroscopy, are laborious and have serious technical
limitations. Single-particle cryogenic electron microscopy (cryo-EM) has many advantages over crystallography
and NMR but is applicable only for large molecules or macromolecular assemblies and cannot be used for the
majority of natural RNAs because of their insufficient size. This proposal is focused on developing a novel
approach for preparing cryo-EM samples of RNA that circumvents the size restrictions, omits the RNA
purification and RNA refolding steps, and allows facile cryo-EM data processing and structure solution. The
proposed proof-of-concept study combines three specific aims. Specific Aim 1 is devoted to the development
of the novel biochemical approach for preparing uniform RNA species by in vitro transcription. Specific Aim 2
will use computational modelling, biochemical assays, and single-particle cryo-EM experimentation to develop
a methodology for preparing cryo-EM samples compatible with the structure solution of small- and medium-
sized RNAs. Aim 3 will validate the methodology using model RNA molecules and conventional single particle
cryo-EM structure solution pipeline. The proposal integrates computational methods, biochemical assays, and
cryo-EM-based structure determination to develop a universal and simple approach for solving structures of
the majority of RNA and RNA-drug complexes. The proposed technology is anticipated to be superior to the
existing methods in labor, cost, and applicability.

## Key facts

- **NIH application ID:** 10910171
- **Project number:** 5R21GM151508-02
- **Recipient organization:** NEW YORK UNIVERSITY SCHOOL OF MEDICINE
- **Principal Investigator:** Alexander Serganov
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $169,500
- **Award type:** 5
- **Project period:** 2023-09-01 → 2026-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10910171

## Citation

> US National Institutes of Health, RePORTER application 10910171, A universal approach for determining three-dimensional RNA structures (5R21GM151508-02). Retrieved via AI Analytics 2026-05-26 from https://api.ai-analytics.org/grant/nih/10910171. Licensed CC0.

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