Construction of a novel phagemid to produce custom DNA origami scaffolds
Construction of a novel phagemid to produce custom DNA origami scaffolds
复制标题
DOI：
10.1093/synbio/ysy015
复制
发表时间：
2018-01-01
影响因子：
3.2
通讯作者：
Douglas, Shawn M.
中科院分区：
文献类型：
--
作者：
Nafisi, Parsa M.;Aksel, Tural;Douglas, Shawn M.
DNA折纸是一种构建纳米级物体的方法，它依赖于一条长的DNA单链作为“支架”，以模板组装许多短的DNA寡核苷酸“订书钉”。生成定制支架序列的能力可以极大地有益于DNA折纸设计过程。定制支架序列可以提供对最终对象的总体大小的更好控制和对低水平结构细节的更好控制，例如对象内特定碱基对的位置。丝状噬菌体和相关的噬菌粒可以很好地作为定制支架DNA的来源。然而，来源于大肠杆菌的支架需要包含多酶DNA序列以便在宿主细菌中生长，并且这些序列不能被改变或去除。这些固定序列区域限制了DNA折纸的设计可能性。在这里，我们报告了一种新的噬菌粒，pScaf，生产支架，有一个定制的序列与一个小得多的393个碱基的固定区域的建设。我们使用pScaf产生新的支架的大小范围从1512到10 080个碱基，并证明了它们在各种DNA折纸形状和组装中的用途。我们预计我们的pScaf噬菌粒将促进DNA折纸方法的发展及其未来的应用。
DNA origami, a method for constructing nanoscale objects, relies on a long single strand of DNA to act as the 'scaffold' to template assembly of numerous short DNA oligonucleotide 'staples'. The ability to generate custom scaffold sequences can greatly benefit DNA origami design processes. Custom scaffold sequences can provide better control of the overall size of the final object and better control of low-level structural details, such as locations of specific base pairs within an object. Filamentous bacteriophages and related phagemids can work well as sources of custom scaffold DNA. However, scaffolds derived from phages require inclusion of multi-kilobase DNA sequences in order to grow in host bacteria, and those sequences cannot be altered or removed. These fixed-sequence regions constrain the design possibilities of DNA origami. Here, we report the construction of a novel phagemid, pScaf, to produce scaffolds that have a custom sequence with a much smaller fixed region of 393 bases. We used pScaf to generate new scaffolds ranging in size from 1512 to 10 080 bases and demonstrated their use in various DNA origami shapes and assemblies. We anticipate our pScaf phagemid will enhance development of the DNA origami method and its future applications.