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A Catalytic DNA Probe with Stem-loop Motif for Human T47D Breast Cancer Cells

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Abstract

In vitro selection methods allow for isolation of DNAzymes (catalytic DNAs) from random DNA pools. Here we describe a fluorogenic DNAzyme, LYF5, isolated using a double-random selection approach: a random DNA pool was selected against a complex molecular mixture derived from a breast cancer cell line, T47D. LYF5 specifically indicates the T47D breast cancer cell line with high sensitivity. After sequence optimization, the second-generation DNAzyme, 2G-LYF5, exhibited an approximately 2-fold higher cleavage percentage. Finally, we have determined that the intramolecular stem-loop motif plays a crucial role in 2G-LYF5 activity. Our findings underscore the capability of singlestranded DNA molecules to perform highly sophisticated functions that are amenable to the development of diagnostic tests for early identification of breast cancer.

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Acknowledgments

The authors thank Mr. Li Y. F. and Mr. Shen Z. F. at the Department of Biochemistry and Biomedical Sciences of McMaster University, Canada, for invaluable guidance in the pre-design and experiment stages. The authors also acknowledge financial support from the Ministry of Science and Technology of China (2012ZX09506001-010), National Natural Science Foundation of China (Nos. 21302108 and 21402105) and Shenzhen Municipal Government SZSITIC (Nos. KC2013ZDZJ0019A, JCYJ20130402145002379, and CXB201104210013A).

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Authors and Affiliations

  1. Tsinghua University, Beijing, 100084, P. R. China

    Yuyang Jiang

  2. The Ministry-Province Jointly Constructed Base for State Key Lab-Shenzhen Key Laboratory of Chemical Biology, the Graduate School at Shenzhen, Tsinghua University, Shenzhen, Guangdong, 518055, P. R. China

    Feng Liu, Jing Zheng & Yuyang Jiang

  3. Department of Pharmacology and Pharmaceutical Sciences, School of Medicine, Tsinghua University, Beijing, 100084, P. R. China

    Yuyang Jiang

  4. Shenzhen Kivita Innovative Drug Discovery Institute, Shenzhen, Guangdong, 518055, P. R. China

    MeiYun Zeng

Authors
  1. Fei Gao
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  2. Feng Liu
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  3. Jing Zheng
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  4. MeiYun Zeng
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  5. Yuyang Jiang
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Correspondence to Feng Liu or Yuyang Jiang.

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Gao, F., Liu, F., Zheng, J. et al. A Catalytic DNA Probe with Stem-loop Motif for Human T47D Breast Cancer Cells. ANAL. SCI. 31, 815–822 (2015). https://doi.org/10.2116/analsci.31.815

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  • DOI: https://doi.org/10.2116/analsci.31.815

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