Fluorescence Resonance Energy Transfer-Based DNA Tetrahedron Nanotweezer for Highly Reliable Detection of Tumor-Related mRNA in Living Cells.
Accurate detection and imaging of tumor-related mRNA in living cells hold great promise for early cancer detection. However, currently, most probes designed to image intracellular mRNA confront intrinsic interferences arising from complex biological matrices and resulting in inevitable false-positive signals. To circumvent this problem, an intracellular DNA nanoprobe, termed DNA tetrahedron nanotweezer (DTNT), was developed to reliably image tumor-related mRNA in living cells based on the FRET (fluorescence resonance energy transfer) "off" to "on" signal readout mode. DTNT was self-assembled from four single-stranded DNAs. In the absence of target mRNA, the respectively labeled donor and acceptor fluorophores are separated, thus inducing low FRET efficiency. However, in the presence of target mRNA, DTNT alters its structure from the open to closed state, thus bringing the dual fluorophores into close proximity for high FRET efficiency. The DTNT exhibited high cellular permeability, fast response and excellent biocompatibility. Moreover, intracellular imaging experiments showed that DTNT could effectively distinguish cancer cells from normal cells and, moreover, distinguish among changes of mRNA expression levels in living cells. The DTNT nanoprobe also exhibits minimal effect of probe concentration, distribution and laser power as other ratiometric probe. More importantly, as a result of the FRET "off" to "on" signal readout mode, the DTNT nanoprobe almost entirely avoids false-positive signals due to intrinsic interferences, such as nuclease digestion, protein binding and thermodynamic fluctuations in complex biological matrices. This design blueprint can be applied to the development of powerful DNA nanomachines for biomedical research and clinical early diagnosis.
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Lei
L
He
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Dan-Qing
DQ
Lu
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Hao
H
Liang
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Sitao
S
Xie
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Can
C
Luo
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Miaomiao
M
Hu
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Liujun
L
Xu
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.
Xiaobing
X
Zhang
Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Life Sciences, and Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University , Changsha 410082, China.|Department of Chemistry and Department of Physiology and Functional Genomics, Center for Research at the Bio/Nano Interface, Health Cancer Center, University of Florida , Gainesville, Florida 32611-7200, United States.
Weihong
0000-0002-8066-1524
W
Tan
D014408:Biomarkers, Tumor / Q000032:analysis; D045744:Cell Line, Tumor; D002470:Cell Survival; D004277:DNA, Single-Stranded / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D057230:Limit of Detection; D049329:Nanostructures / Q000737:chemistry*; D009369:Neoplasms / Q000737:chemistry* / Q000175:diagnosis; D061848:Optical Imaging / Q000379:methods; D012333:RNA, Messenger / Q000032:analysis*; D013816:Thermodynamics
D014408:Biomarkers, Tumor; D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D012333:RNA, Messenger
DNA tetrahedron; fluorescence resonance energy transfer; fluorescent imaging; self-assembly; tumor-related mRNA