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| Title | A generalizable scaffold-based approach for structure determination of RNAs by cryo-EM. |
|---|---|
| Journal, issue, pages | Nucleic Acids Res, Vol. 51, Issue 20, Page e100, Year 2023 |
| Publish date | Nov 10, 2023 |
 Authors | Conner J Langeberg / Jeffrey S Kieft /  |
| PubMed Abstract | Single-particle cryo-electron microscopy (cryo-EM) can reveal the structures of large and often dynamic molecules, but smaller biomolecules (≤50 kDa) remain challenging targets due to their ...Single-particle cryo-electron microscopy (cryo-EM) can reveal the structures of large and often dynamic molecules, but smaller biomolecules (≤50 kDa) remain challenging targets due to their intrinsic low signal to noise ratio. Methods to help resolve small proteins have been applied but development of similar approaches to aid in structural determination of small, structured RNA elements have lagged. Here, we present a scaffold-based approach that we used to recover maps of sub-25 kDa RNA domains to 4.5-5.0 Å. While lacking the detail of true high-resolution maps, these maps are suitable for model building and preliminary structure determination. We demonstrate this method helped faithfully recover the structure of several RNA elements of known structure, and that it promises to be generalized to other RNAs without disturbing their native fold. This approach may streamline the sample preparation process and reduce the optimization required for data collection. This first-generation scaffold approach provides a robust system to aid in RNA structure determination by cryo-EM and lays the groundwork for further scaffold optimization to achieve higher resolution. |
 External links | Nucleic Acids Res / PubMed:37791881 / PubMed Central |
| Methods | EM (single particle) |
| Resolution | 2.44 - 3.46 Å |
| Structure data | EMDB-41308, PDB-8tjq: EMDB-41311, PDB-8tju: EMDB-41312, PDB-8tjv: EMDB-41313, PDB-8tjx: |
| Chemicals |  ChemComp-MG:  ChemComp-F:  ChemComp-K: |
| Source |
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 Keywords |  RNA / Ribozyme / xrRNA / Scaffold |
