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Selection and characterization of a DNA aptamer to crystal violet

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Abstract

Aptamers are short single-stranded DNA or RNA, which can be selected in vitro by systematic evolution of ligands by exponential enrichment (SELEX). In order to develop novel light-up probes to substitute G-quadruplex (G4), we selected a DNA aptamer for crystal violet (CV), a triphenylmethane light-up dye, by a modified affinity chromatography-based SELEX. The ssDNA pool was first coupled on streptavidin-coated agarose beads through a biotin labeled complementary oligonucleotide, and then the aptamer sequences would be released from agarose beads by CV affinity. This method is simple, straightforward and effective. The aptamer sequence with a low micromolar dissociation constant (Kd) and good specificity was achieved after 11 rounds of selection. The light-up properties of the CV–aptamer were also investigated, and the CV showed dramatic fluorescence enhancement. The CV–aptamer pair could be further used as a novel light-up fluorescent probe to design biosensors.

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Acknowledgements

This work was supported by the National Natural Science Foundation of China (No. 21575154, 21775160, 21507156) and the Science and Technology Foundation of Suzhou (No. SYG201526).

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Authors and Affiliations

  1. CAS Key Laboratory of Nano-Bio Interface, Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, Suzhou, 215123, China

    Yang Chen, Jine Wang, Yajie Zhang, Lijun Xu, Tian Gao & Renjun Pei

  2. School of Pharmacy, Xi’an Jiaotong University, Xi’an, 710061, China

    Yang Chen & Bing Wang

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  1. Yang Chen
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  2. Jine Wang
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  3. Yajie Zhang
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  5. Tian Gao
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  7. Renjun Pei
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Correspondence to Renjun Pei.

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Chen, Y., Wang, J., Zhang, Y. et al. Selection and characterization of a DNA aptamer to crystal violet. Photochem Photobiol Sci 17, 800–806 (2018). https://doi.org/10.1039/c7pp00457e

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