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. 2023:2568:243-249.
doi: 10.1007/978-1-0716-2687-0_16.

Mix-and-Inject Serial Femtosecond Crystallography to Capture RNA Riboswitch Intermediates

Jason R Stagno  1 , Juraj Knoska  2   3 , Henry N Chapman  2   3   4 , Yun-Xing Wang  5
Affiliations

Mix-and-Inject Serial Femtosecond Crystallography to Capture RNA Riboswitch Intermediates

Jason R Stagno et al. Methods Mol Biol. 2023.

Abstract

Time-resolved structure determination of macromolecular conformations and ligand-bound intermediates is extremely challenging, particularly for RNA. With rapid technological advances in both microfluidic liquid injection and X-ray free electron lasers (XFEL), a new frontier has emerged in time-resolved crystallography whereby crystals can be mixed with ligand and then probed with X-rays (mix-and-inject) in real time and at room temperature. This chapter outlines the basic setup and procedures for mix-and-inject experiments for recording time-resolved crystallographic data of riboswitch RNA reaction states using serial femtosecond crystallography (SFX) and an XFEL.

Keywords: Diffusive mixing; RNA conformational intermediates; Serial femtosecond crystallography; Time-resolved crystallography; X-ray free electron laser.

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