Cryo-EM for DNA Samples

Cryo-electron microscopy (cryo-EM) can be used to visualize DNA or RNA structures based on our cryo-EM platform provided by Creative Biostructure. We develop a novel suite of tools to overcome the size and throughput limitations of cryo-EM, which can be applied to small molecules such as DNA or RNA. It allows us to create soluble nanostructures with an unprecedented combination of spatial resolution and chemical versatility. In principle, we can attach any moiety to our devices as long as it has a DNA binding domain or it can be coupled to DNA.

Cryo-EM of vitrified specimens became a practical method and opened the possibility of observing biomolecules in their aqueous environment. This technology made it possible to omit chemical fixation and staining. Most important, it allowed a much better preservation of the spatial arrangement. Using conventional cryo-EM technology for large biomolecules (large proteins, membrane proteins, VLPs), it allows us to image hundreds of thousands of single molecules frozen in a single molecule thick layer of vitreous ice. From these images, we then use algorithms to reconstruct the molecular structure of the protein in multiple conformations. Nowadays, Cryo-EM is transforming structural biology allowing us to study many various targets for the first time and gain new mechanistic insights to help guide future drug design.

Our research team has experience in exploiting cryo-EM image analysis to obtain structural information to reveal the configuration of a designed DNA nanostructure (Figure 1). We successfully obtained the 3D classification model of the DNA samples offered by our customer. All the 2D TEM particles were classified by the 3D similarity and consistence by using 3D classification function. For practical applications, it is essential to verify that the 3D structures are accurately produced as designed, which requires high resolution, at least sufficient to resolve the DNA helix.

Figure 1. 3D classification of DNA sample using cryo-EM

Cryo-EM is transforming structural biology allowing us to study many target proteins for the first time and gain new mechanistic insights to help guide future drug design. Creative Biostructure has launched this technology to visualize small molecules such as DNA or RNA. Having access to this technology through the combination of our unique nanopoarticles and cryo-EM platform really helps keep Creative Biostructure at the leading edge in the industry. Cryo-EM services have become our routine work, and part of an organization investing, collaborating with the best research groups in the world, from both industry and academia. To position it for the first time within the drug discovery setting is really exciting!

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Reference

1. Takayuki Kato, Russell P. Goodman et al. (2009) High-Resolution Structural Analysis of a DNA Nanostructure by cryoEM. Nano Lett. 2009, 9 (7), pp 2747–2750.