Engineering Micrometer‐Sized DNA Tracks for High‐Speed DNA Synthesis and Biosensing. Issue 51 (9th November 2020)

• Record Type: Journal Article
• Authors: Wang, Liying; Song, Kaiyun; Qu, Yuanyuan; Chang, Yangyang; Li, Zhongping; Dong, Chuan; Liu, Meng; Brennan, John D.; Li, Yingfu
• Is Part Of: Angewandte Chemie international edition. Volume 59:Issue 51(2020)
• Page Start: 22947
• Page End: 22951
• Publication Date: 2020-11-09
• Subjects: biosensors -- polymerase -- rolling circle amplification -- nucleic acids -- self-assembly; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.202010693
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: φ 29 DNA polymerase (Pol φ 29) is capable of synthesizing long‐chain single‐stranded (ss) DNA molecules by copying the sequence of a small ss circular DNA template (ssCDT) in a process known as rolling circle amplification (RCA). The use of a ssCDT in RCA, however, comes with a key drawback: the rate of DNA synthesis is significantly reduced. We hypothesize that this issue can be overcome using a very long linear ssDNA template with a repeating sequence. To test this idea, we engineered a DNA assembly, which we denote "micrometer‐sized DNA track" (μDT). This μDT, with an average length of ≈13.5 μm, is made of a long chain DNA with a primer‐binding domain at its 3′ end and ≈1000 repeating sequence units at its 5′ end, each carrying a DNA anchor. We find that Pol φ 29 copies μDT at a speed ≈5‐time faster than it does a related ssCDT. We use this to design a simple all‐in‐one printed paper device for rapid and sensitive detection of microRNA let‐7. This paper sensor is capable of detecting 1 pM let‐7a in 10 minutes. Abstract : A micrometer‐sized DNA track containing one primer‐binding domain at the 3′ end and ≈1000 repeated DNA anchorages at the 5′ end was engineered in a nearly linear fashion, which serves as an efficient template for in vitro DNA synthesis by polymerase at a maximum speed and processivity. This was used to produce a paper‐based microRNA sensor capable of detecting 1 pM let‐7a in 10 minutes.