期刊
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
卷 54, 期 33, 页码 9637-9641出版社
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201503182
关键词
aptamers; biosensors; DNA amplification; rolling circle amplification; structure switching
资金
- Natural Sciences and Engineering Research Council of Canada (NSERC)
- Pro-Lab Diagnostics Inc
- Ontario Ministry of Research and Innovation
- Canada Foundation for Innovation (CFI)
phi 29 DNA polymerase (phi 29DP) is able to carry out repetitive rounds of DNA synthesis using a circular DNA template by rolling circle amplification (RCA). It also has the ability to execute 3-5 digestion of single-stranded but not double-stranded DNA. A biosensor engineering strategy is presented that takes advantage of these two properties of phi 29DP coupled with structure-switching DNA aptamers. The design employs a DNA assembly made of a circular DNA template, a DNA aptamer, and a pre-primer. The DNA assembly is unable to undergo RCA in the absence of cognate target owing to the formation of duplex structures. The presence of the target, however, triggers a structure-switching event that causes nucleolytic conversion of the pre-primer by phi 29DP into a mature primer to facilitate RCA. This method relays target detection by the aptamer to the production of massive DNA amplicons, giving rise to dramatically enhanced detection sensitivity.
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