Precise Coating of a Wide Range of DNA Templates by a Protein Polymer with a DNA Binding Domain

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• 作者：Armando Hernandez-GarciaNicole A. Estrich ; Marc W. T. Werten ; Johan R. C. Van Der Maarel ; Thomas H. LaBean ; Frits A. de WolfMartien A. Cohen Stuart ; Renko de Vries
• 刊名：ACS Nano
• 出版年：2017
• 出版时间：January 24, 2017
• 年：2017
• 卷：11
• 期：1
• 页码：144-152
• 全文大小：520K
• ISSN：1936-086X

文摘

Emerging DNA-based nanotechnologies would benefit from the ability to modulate the properties (e.g., solubility, melting temperature, chemical stability) of diverse DNA templates (single molecules or origami nanostructures) through controlled, self-assembling coatings. We here introduce a DNA coating agent, called C₈–B^Sso7d, which binds to and coats with high specificity and affinity, individual DNA molecules as well as folded origami nanostructures. C₈–B^Sso7d coats and protects without condensing, collapsing or destroying the spatial structure of the underlying DNA template. C₈–B^Sso7d combines the specific nonelectrostatic DNA binding affinity of an archeal-derived DNA binding domain (Sso7d, 7 kDa) with a long hydrophilic random coil polypeptide (C₈, 73 kDa), which provides colloidal stability (solubility) through formation of polymer brushes around the DNA templates. C₈–B^Sso7d is produced recombinantly in yeast and has a precise (but engineerable) amino acid sequence of precise length. Using electrophoresis, AFM, and fluorescence microscopy we demonstrate protein coat formation with stiffening of one-dimensional templates (linear dsDNA, supercoiled dsDNA and circular ssDNA), as well as coat formation without any structural distortion or disruption of two-dimensional DNA origami template. Combining the programmability of DNA with the nonperturbing precise coating capability of the engineered protein C₈–B^Sso7d holds promise for future applications such as the creation of DNA–protein hybrid networks, or the efficient transfection of individual DNA nanostructures into cells.