@Article{Liu2022,
author={Liu, Di
and Th{\'e}lot, Fran{\c{c}}ois A.
and Piccirilli, Joseph A.
and Liao, Maofu
and Yin, Peng},
title={Sub-3-{\AA} cryo-EM structure of RNA enabled by engineered homomeric self-assembly},
journal={Nature Methods},
year={2022},
month={May},
day={02},
abstract={High-resolution structural studies are essential for understanding the folding and function of diverse RNAs. Herein, we present a nanoarchitectural engineering strategy for efficient structural determination of RNA-only structures using single-particle cryogenic electron microscopy (cryo-EM). This strategy---ROCK (RNA oligomerization-enabled cryo-EM via installing kissing loops)---involves installing kissing-loop sequences onto the functionally nonessential stems of RNAs for homomeric self-assembly into closed rings with multiplied molecular weights and mitigated structural flexibility. ROCK enables cryo-EM reconstruction of the Tetrahymena group I intron at 2.98-{\AA} resolution overall (2.85{\thinspace}{\AA} for the core), allowing de novo model building of the complete RNA, including the previously unknown peripheral domains. ROCK is further applied to two smaller RNAs---the Azoarcus group I intron and the FMN riboswitch, revealing the conformational change of the former and the bound ligand in the latter. ROCK holds promise to greatly facilitate the use of cryo-EM in RNA structural studies.},
issn={1548-7105},
doi={10.1038/s41592-022-01455-w},
url={https://doi.org/10.1038/s41592-022-01455-w}
}