Near-Atomic Resolution Cryo-EM Image Reconstruction of RNA

Shanshan Li; Kaiming Zhang; Wah Chiu

doi:10.1007/978-1-0716-2687-0_12

Near-Atomic Resolution Cryo-EM Image Reconstruction of RNA

Methods Mol Biol. 2023:2568:179-192. doi: 10.1007/978-1-0716-2687-0_12.

Authors

Shanshan Li^{1

2}, Kaiming Zhang^{1

2}, Wah Chiu^{3

4}

Affiliations

¹ MOE Key Laboratory for Cellular Dynamics and Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, China.
² Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA, USA.
³ Department of Bioengineering, James H. Clark Center, Stanford University, Stanford, CA, USA. wahc@stanford.edu.
⁴ CryoEM and Bioimaging Division, Stanford Synchrotron Radiation Lightsource, SLAC National Accelerator Laboratory, Stanford University, Menlo Park, CA, USA. wahc@stanford.edu.

PMID: 36227569
DOI: 10.1007/978-1-0716-2687-0_12

Abstract

The rapid development of cryogenic electron microscopy (cryo-EM) enables the structure determination of macromolecules without the need for crystallization. Protein, protein-lipid, and protein-nucleic acid complexes can now be routinely resolved by cryo-EM single-particle analysis (SPA) to near-atomic or atomic resolution. Here we describe the structure determination of pure RNAs by SPA, from cryo-specimen preparation to data collection and 3D reconstruction. This protocol is useful to yield many cryo-EM structures of RNA, here exemplified by the Tetrahymena L-21 ScaI ribozyme at 3.1-Å resolution.

Keywords: Cryo-EM; Protocol; RNA; Ribozyme; Single-particle analysis.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Cryoelectron Microscopy / methods
Image Processing, Computer-Assisted / methods
Lipids
Proteins
RNA*
RNA, Catalytic*

Substances

Lipids
Proteins
RNA, Catalytic
RNA

Abstract

Publication types

MeSH terms

Substances

Grants and funding