Simultaneous Enzyme-Free Detection of Multiple Long Noncoding RNAs in Cancer Cells at Single-Molecule/Particle Level

Aberrant change in long noncoding RNA (lncRNA) is associated with various diseases and cancers. So far, simultaneous detection of lncRNAs has remained a great challenge due to their large size and extensive secondary structure. Herein, we develop an enzyme-free single-molecule/particle detection met...

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Published inNano letters Vol. 21; no. 10; pp. 4193 - 4201
Main Authors Zhang, Yan, Wang, Chen, Zou, Xiaoran, Tian, Xiaorui, Hu, Juan, Zhang, Chun-yang
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 26.05.2021
Subjects
long noncoding RNA (lncRNA)
cancer diagnosis
single-molecule/particle detection
living cell imaging
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Abstract Aberrant change in long noncoding RNA (lncRNA) is associated with various diseases and cancers. So far, simultaneous detection of lncRNAs has remained a great challenge due to their large size and extensive secondary structure. Herein, we develop an enzyme-free single-molecule/particle detection method for simultaneous detection of multiple lncRNAs in cancer cells based on target-catalyzed strand displacement. We designed the magnetic bead–capture probe–multiple Cy5/Cy3-modified reporter unit complexes to isolate and identify lncRNA MALAT1 and lncRNA HOTAIR. The target-catalyzed strand displacement reactions lead to the release of Cy5 and Cy3 fluorescent molecules from the complexes, which can be subsequently quantified by single-molecule/particle detection. The dual-targetability, good selectivity and high sensitivity of this method enables simultaneous detection of multiple lncRNAs in even single cancer cell. Importantly, this method can discriminate cancer cells from normal cells and has significant advantages in the simple sequence design and in being free of enzymes, holding great potential in living cell imaging and early clinical diagnosis.
AbstractList Aberrant change in long noncoding RNA (lncRNA) is associated with various diseases and cancers. So far, simultaneous detection of lncRNAs has remained a great challenge due to their large size and extensive secondary structure. Herein, we develop an enzyme-free single-molecule/particle detection method for simultaneous detection of multiple lncRNAs in cancer cells based on target-catalyzed strand displacement. We designed the magnetic bead-capture probe-multiple Cy5/Cy3-modified reporter unit complexes to isolate and identify lncRNA MALAT1 and lncRNA HOTAIR. The target-catalyzed strand displacement reactions lead to the release of Cy5 and Cy3 fluorescent molecules from the complexes, which can be subsequently quantified by single-molecule/particle detection. The dual-targetability, good selectivity and high sensitivity of this method enables simultaneous detection of multiple lncRNAs in even single cancer cell. Importantly, this method can discriminate cancer cells from normal cells and has significant advantages in the simple sequence design and in being free of enzymes, holding great potential in living cell imaging and early clinical diagnosis.
Author Hu, Juan
Zhang, Yan
Wang, Chen
Zhang, Chun-yang
Tian, Xiaorui
Zou, Xiaoran
AuthorAffiliation School of Chemistry and Chemical Engineering
College of Chemistry, Chemical Engineering and Materials Science, Collaborative Innovation Center of Functionalized Probes for Chemical Imaging in Universities of Shandong, Key Laboratory of Molecular and Nano Probes, Ministry of Education, Shandong Provincial Key Laboratory of Clean Production of Fine Chemicals
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cancer diagnosis
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Snippet Aberrant change in long noncoding RNA (lncRNA) is associated with various diseases and cancers. So far, simultaneous detection of lncRNAs has remained a great...
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Title Simultaneous Enzyme-Free Detection of Multiple Long Noncoding RNAs in Cancer Cells at Single-Molecule/Particle Level
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