期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 59, 期 51, 页码 22947-22951出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.202010693
关键词
biosensors; polymerase; rolling circle amplification; nucleic acids; self-assembly
资金
- National Natural Science Foundation of China (NSFC) [21922601, 21777013]
- Recruitment Program of Global Young Experts (Thousand Talents Program)
- LiaoNing Revitalization Talents Program [XLYC1807080]
- Fundamental Research Funds for the Central Universities [DUT19TD20]
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- Canadian Foundation for Innovation (CFI)
- Canadian Institutes of Health Research (CIHR)
- Ontario Ministry for Research and Innovation [RE07-045]
phi 29 DNA polymerase (Pol phi 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 (mu DT). This mu DT, with an average length of approximate to 13.5 mu m, is made of a long chain DNA with a primer-binding domain at its 3 ' end and approximate to 1000 repeating sequence units at its 5 ' end, each carrying a DNA anchor. We find that Pol phi 29 copies mu DT at a speed approximate to 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.
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