Abstract
BackgroundPost-crystallization dehydration methods, applying either vapor diffusion or humidity control devices, have been widely used to improve the diffraction quality of protein crystals. Despite the fact that RNA crystals tend to diffract poorly, there is a dearth of reports on the application of dehydration methods to improve the diffraction quality of RNA crystals.ResultsWe use dehydration techniques with a Free Mounting System (FMS, a humidity control device) to recover the poor diffraction quality of RNA crystals. These approaches were applied to RNA constructs that model various RNA-mediated repeat expansion disorders.ConclusionThe method we describe herein could serve as a general tool to improve diffraction quality of RNA crystals to facilitate structure determinations.Electronic supplementary materialThe online version of this article (doi:10.1186/s12900-016-0069-1) contains supplementary material, which is available to authorized users.
Highlights
Post-crystallization dehydration methods, applying either vapor diffusion or humidity control devices, have been widely used to improve the diffraction quality of protein crystals
Diffraction images of the crystals were collected every 5 min, while the relative humidity (Rh) of the crystals was reduced to 70 % at a gradient of 0.25 % Rh change per min
Construct design and crystallization To overcome the inherent limitation of intermolecular crystal contact in RNAs, the GAAA tetraloop and the tetraloop receptor have been utilized as a general module to promote RNA crystallization [18, 19]
Summary
Post-crystallization dehydration methods, applying either vapor diffusion or humidity control devices, have been widely used to improve the diffraction quality of protein crystals. Despite the fact that RNA crystals tend to diffract poorly, there is a dearth of reports on the application of dehydration methods to improve the diffraction quality of RNA crystals. Results: We use dehydration techniques with a Free Mounting System (FMS, a humidity control device) to recover the poor diffraction quality of RNA crystals. These approaches were applied to RNA constructs that model various RNA-mediated repeat expansion disorders. Conclusion: The method we describe could serve as a general tool to improve diffraction quality of RNA crystals to facilitate structure determinations. A common method to overcome this problem is to re-design the construct and repeat the crystallization screening until a perfect construct is obtained for diffraction experiments [1, 2]
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