A light-up fluorescence assay for tumor cell detection based on bifunctional split aptamers

The Analyst
Yuqiong SunKemin Wang

Abstract

Light-up aptamers have attracted growing attention due to their advantages of being label-free and having low fluorescence background. In this work, we developed a light-up fluorescence assay for label-free detection of tumor cells based on a bifunctional split aptamer (BFSA) that contained two DNA strands (BFSA-a and BFSA-b). BFSA-a and BFSA-b were constructed by combining aptamers ZY11 and ThT.2-2, which could specifically bind to the tumor cell SMMC-7721 and activate the fluorescence of thioflavin T (ThT). A Helper strand was introduced to hybridize with BFSA-b, and then BFSA-a and BFSA-b were separated if the target cell was absent. Only when the target cell is present can BFSA-a approach and hybridize with BFSA-b due to the 'induced-fit effect', which made the Helper strand dissociate. Then ThT bound to BFSA and the fluorescence of ThT was activated. The results indicated that this fluorescence assay had a good linear response to the target cells in the range of 250-20 000 cells in 100 μL binding buffer; the lowest cell number actually detected was 125 cells in 100 μL buffer. This assay also displayed excellent selectivity and was successfully applied to detect target cells in 20% human serum samples. The design of bifunct...Continue Reading

References

May 26, 1999·Proceedings of the National Academy of Sciences of the United States of America·D Grate, C Wilson
Dec 4, 2003·Journal of the American Chemical Society·Jeremy R BabendureRoger Y Tsien
Mar 21, 2007·Chembiochem : a European Journal of Chemical Biology·Dihua ShangguanWeihong Tan
Jun 12, 2010·Nature Protocols·Kwame SefahWeihong Tan
Jul 2, 2010·Nature Reviews. Drug Discovery·Anthony D KeefeAndrew Ellington
Sep 18, 2012·Protein & Cell·Tianjiao Wang, Judhajeet Ray
Oct 30, 2014·Analytical Chemistry·Chengzhou ZhuYuehe Lin
Feb 13, 2015·Chemical Communications : Chem Comm·Qing LiJian-Hui Jiang
Dec 15, 2015·Chemical Communications : Chem Comm·Baoyin YuanXiayu Zhao
Jan 23, 2016·Chemical Communications : Chem Comm·Fei XuHong-Yuan Chen
Feb 13, 2016·The Analyst·Muslum Ilgu, Marit Nilsen-Hamilton
Feb 19, 2016·Chemical Communications : Chem Comm·Teru KatoMasumi Hyuga
Mar 10, 2016·Chemical Society Reviews·Hong-Min MengXiao-Bing Zhang
May 14, 2016·The Analyst·Chunmei GuHanchang Shi
May 14, 2016·Analytical and Bioanalytical Chemistry·Shunxiang GaoLianghua Wang
Jul 26, 2016·Frontiers in Chemistry·Jonathan Ouellet
Nov 4, 2016·Nature Reviews. Drug Discovery·Jiehua Zhou, John Rossi
Feb 9, 2017·Nanoscale Research Letters·Zixi HuXiaoling Lu
Jun 22, 2017·Journal of the American Chemical Society·Hailan KuaiWeihong Tan
Jul 18, 2017·Journal of the American Chemical Society·Zhan-Ming YingJian-Hui Jiang
Nov 22, 2000·Journal of the American Chemical Society·Milan N StojanovicDonald W Landry
Jan 4, 2018·International Journal of Molecular Sciences·Farah BouheddaMichael Ryckelynck
May 18, 2018·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Yang ChenRenjun Pei

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Citations

Nov 9, 2019·Chembiochem : a European Journal of Chemical Biology·Sotaro MisuKazuhito Tanabe
Mar 1, 2020·Nucleic Acids Research·Mégane DebiaisSabine Müller
Mar 26, 2020·Angewandte Chemie·Dmitry M Kolpashchikov, Alexander A Spelkov
Nov 5, 2020·Analytical Chemistry·Andresa B BezerraChristopher J Easley

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Methods Mentioned

BETA
flow cytometry
fluorescence assay

Software Mentioned

BFSA
Helper

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