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ABSTRACT The development of high-throughput RNA structure profiling methods in the past decade has greatly facilitated our ability to map and characterize different aspects of RNA structures

transcriptome-wide in cell populations, single cells and single molecules. The resulting high-resolution data have provided insights into the static and dynamic nature of RNA structures,

revealing their complexity as they perform their respective functions in the cell. In this Review, we discuss recent technical advances in the determination of RNA structures, and the roles

of RNA structures in RNA biogenesis and functions, including in transcription, processing, translation, degradation, localization and RNA structure-dependent condensates. We also discuss the

current understanding of how RNA structures could guide drug design for treating genetic diseases and battling pathogenic viruses, and highlight existing challenges and future directions in

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SIMILAR CONTENT BEING VIEWED BY OTHERS RNA STRUCTURE PROBING UNCOVERS RNA STRUCTURE-DEPENDENT BIOLOGICAL FUNCTIONS Article 25 June 2021 PROBING THE DYNAMIC RNA STRUCTUROME AND ITS FUNCTIONS

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distribution. _RNA_ 23, 995–1011 (2017). Article  CAS  PubMed  PubMed Central  Google Scholar  Download references ACKNOWLEDGEMENTS This work was supported by the UK Biotechnology and

Biological Sciences Research Council (BBSRC) (BB/X01102X/1) and European Research Council (ERC) (selected by the ERC, funded by BBSRC Horizon Europe Guarantee (EP/Y009886/1)) (Y.D. and

Y.Z.). Y.W. and X.C. are supported by funding from A*STAR, the National Research Foundation of Singapore, the EMBO Young Investigator Programme and a CIFAR Azrieli global scholar fellowship.

AUTHOR INFORMATION Author notes * These authors contributed equally: Xinang Cao, Yueying Zhang. AUTHORS AND AFFILIATIONS * Stem Cell and Regenerative Biology, Genome Institute of Singapore,

Singapore, Singapore Xinang Cao & Yue Wan * Department of Cell and Developmental Biology, John Innes Centre, Norwich, UK Yueying Zhang & Yiliang Ding * Department of Biochemistry,

Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore Yue Wan Authors * Xinang Cao View author publications You can also search for this author inPubMed 

Google Scholar * Yueying Zhang View author publications You can also search for this author inPubMed Google Scholar * Yiliang Ding View author publications You can also search for this

author inPubMed Google Scholar * Yue Wan View author publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS The authors contributed equally to all aspects of

the article. CORRESPONDING AUTHORS Correspondence to Yiliang Ding or Yue Wan. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing interests. PEER REVIEW PEER REVIEW

INFORMATION _Nature Reviews Molecular Cell Biology_ thanks Tetsuro Hirose, Yuanchao Xue and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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SUPPLEMENTARY INFORMATION GLOSSARY * Aptamer domain An RNA structure in a riboswitch that binds to small molecules. * Enzymatic structure probing Refers to the use of nucleases that cleave

RNA selectively at single-stranded or double-stranded regions (for example, RNase T1 and RNase V1, respectively); the resulting digestion footprints of the RNA can chart its structure. *

i-Motifs Cytosine-rich DNAs that form quadruplex structures; also known as intercalated-motif DNAs. * Riboswitches Highly folded segments of (mostly bacterial) mRNAs that, when bound by

environmental small molecules, induce structure changes that regulate the transcription or translation of the mRNA. * R-loop structure A three-stranded nucleic acid structure composed of a

DNA–RNA hybrid and a displaced single strand of DNA. * Small nucleolar RNAs A class of small non-coding RNAs (ncRNAs) that mostly reside in nucleoli, which guide chemical modifications of

other RNA species such as ribosomal RNAs. * Stress granules Dynamic cytoplasmic bodies formed in response to cellular stress, comprising RNA molecules and various proteins; they have a role

in RNA metabolism and are associated with responses to environmental stresses. * Upstream open reading frames (uORFs). Open reading frames (ORFs) located upstream of a main open reading

frame, that is, within the 5′ untranslated region (UTR) of the mRNA. uORFs can encode small peptides, and can regulate the translation of the main ORF by competing for the translation

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permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Cao, X., Zhang, Y., Ding, Y. _et al._ Identification of RNA structures and their roles in RNA functions. _Nat Rev Mol Cell Biol_ 25, 784–801

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