Goodman Brian S, Reck-Peterson Samara L
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.
Department of Cell Biology, Harvard Medical School, Boston, Massachusetts, USA.
Methods Enzymol. 2014;540:169-88. doi: 10.1016/B978-0-12-397924-7.00010-8.
Many cytoskeletal motors function in groups to coordinate the spatial and temporal positioning of cellular cargo. While methods to study the biophysical properties of single motors are well established, methods to understand how multiple motors work synergistically or antagonistically are less well developed. Here, we describe a three-dimensional synthetic cargo structure made using DNA origami, which can be used to template defined numbers and types of cytoskeletal motors with programmable geometries and spacing. We describe methods for building the DNA origami structure, covalently attaching motors to DNA, forming the motor-DNA origami structure complex, and single-molecule assays to examine the motile properties of motor ensembles.
许多细胞骨架马达协同发挥作用，以协调细胞货物的空间和时间定位。虽然研究单个马达生物物理特性的方法已经很成熟，但了解多个马达如何协同或拮抗工作的方法却不太完善。在这里，我们描述了一种使用DNA折纸制成的三维合成货物结构，它可用于以可编程的几何形状和间距为细胞骨架马达的特定数量和类型提供模板。我们描述了构建DNA折纸结构、将马达共价连接到DNA、形成马达-DNA折纸结构复合物以及用于检查马达组合运动特性的单分子检测方法。
