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Nanopore-based detection of circulating microRNAs in lung cancer patients

Nature Nanotechnology volume 6, pages 668–674 (2011)Cite this article

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Abstract

MicroRNAs are short RNA molecules that regulate gene expression, and have been investigated as potential biomarkers because their expression levels are correlated with various diseases. However, detecting microRNAs in the bloodstream remains difficult because current methods are not sufficiently selective or sensitive. Here, we show that a nanopore sensor based on the α-haemolysin protein can selectively detect microRNAs at the single molecular level in plasma samples from lung cancer patients without the need for labels or amplification of the microRNA. The sensor, which uses a programmable oligonucleotide probe to generate a target-specific signature signal, can quantify subpicomolar levels of cancer-associated microRNAs and can distinguish single-nucleotide differences between microRNA family members. This approach is potentially useful for quantitative microRNA detection, the discovery of disease markers and non-invasive early diagnosis of cancer.

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Figure 1: Capturing single microRNA molecules in the nanopore.
Figure 2: Optimizing the probe sequence for enhanced detection sensitivity.
Figure 3: Differentiation of let-7 microRNAs that contain one or two different nucleotides.
Figure 4: Detection of miR-155 in the plasma of lung cancer patients.

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Acknowledgements

The authors thank K. Gillis, T-C. Hwang, S-J. Chen, F. Hsieh and M. Milanick for invaluable discussions on experimental design and data analysis. This investigation was partially supported by grants from the National Science Foundation 0546165 (L-Q.G.), the National Institutes of Health GM079613 (L-Q.G.) and the University of Missouri Intellectual Property Fast Track Initiative (A8881, M.X.W.), and was conducted in a facility that was constructed with support from the Research Facilities Improvement Program grant no. C06-RR-016489-01 from the National Centre for Research Resources, National Institutes of Health.

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  1. Yong Wang and Dali Zheng: These authors contributed equally to this work

Authors and Affiliations

  1. Department of Biological Engineering and Dalton Cardiovascular Research Center, University of Missouri, Columbia, MO 65211

    Yong Wang, Qiulin Tan & Li-Qun Gu

  2. Ellis Fischel Cancer Center and Department of Pathology and Anatomical Sciences, University of Missouri, Columbia, MO 65211, USA

    Dali Zheng & Michael X. Wang

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  1. Yong Wang
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Contributions

Y.W. designed and performed the nanopore experiments, collected and analysed the nanopore data, and co-wrote the manuscript. D.Z. designed and performed the qRT-PCR experiments, analysed the qRT-PCR data and co-wrote the manuscript. Q.T. performed molecular biology experiments, including protein synthesis. M.X.W. conceived the qRT-PCR experiments, provided the patients' samples and co-wrote the manuscript. L-Q.G. conceived the principal idea, designed the nanopore experiments and wrote the manuscript. All the authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Michael X. Wang or Li-Qun Gu.

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Wang, Y., Zheng, D., Tan, Q. et al. Nanopore-based detection of circulating microRNAs in lung cancer patients. Nature Nanotech 6, 668–674 (2011). https://doi.org/10.1038/nnano.2011.147

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