Controlled cryo-EM sample preparation through DNA-Origami pillars

Objective

Electron cryomicroscopy (cryo-EM) is rapidly emerging as a powerful technique for high-resolution structure determination of protein complexes. Despite recent advances, there is still ample room for improvement of this method, in particular regarding sample preparation. Outstanding challenges are reproducibility of freezing conditions to obtain an optimal ice thickness; to prevent unfolding or preferential orientation of proteins against the hydrophobic air-water interface; and a limited accuracy of beam-induced motion correction for relatively small particles. Here, we propose to study how DNA origami objects that are added in with the sample may affect cryo-EM grid preparation. In addition, by examining a variety of DNA origami structures in the electron microscope, we also aim to identify and improve on structural weaknesses in their design. We will test new design strategies, analyse those, and use them for application in cryo-EM. Thereby, this proposal will facilitate both cryo-EM structure determination and the design of nano-tools that are made from DNA.

Fields of science

• engineering and technology › materials engineering › coating and films
• natural sciences › biological sciences › genetics and heredity › dna
• natural sciences › biological sciences › biochemistry › biomolecules › proteins
• engineering and technology › nanotechnology › nano-materials › two-dimensional nanostructures › graphene

Programme(s)

• H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility

Topic(s)

• MSCA-IF-2014-EF - Marie Skłodowska-Curie Individual Fellowships (IF-EF)

Call for proposal

H2020-MSCA-IF-2014
See other projects for this call

Funding Scheme

MSCA-IF-EF-ST - Standard EF

Coordinator

MEDICAL RESEARCH COUNCIL

Address

North Star Avenue Polaris House 2 Floor David Phillips Building
SN2 1FL Swindon
United Kingdom

Activity type

Research Organisations

EU contribution

€ 183 454,80