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ChemComm
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COMMUNICATION
Journal Name
Discipline Development and Talent Training in Zhejiang
University. J. L. thanks the Thousand Young Talents Plan of
China and Hundred Talents Program of Zhejiang University.
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DOI: 10.1039/C9CC03923F
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Conflicts of interest
There are no conflicts to declare.
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Notes and references
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Fig. 4 Time course study of RNA decay in HeLa cells using p A-labeled and SO
3
-TPE-N
3
-
2
6
conjugated strategy. A-L) Cells which were pretreated with p A (1 mM) for 6 h, were
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incubated with actinomycin D for 0, 2, 4, and 6 h, and then stained with SO
using alkyne-azide click chemistry. Blue channel: excitation wavelength: 405 nm;
collection wavelength: 420-440 nm; yellow channel: excitation wavelength: 405 nm;
3 3
-TPE-N
1
08, 325,
collection wavelength: 520-560 nm. The scale bar is 20 μm. M) Detection of whole cell
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5 H. Kobayashi, M. Ogawa, R. Alford, P. L. Choyke and Y. Urano,
Chem. Rev., 2010, 100, 2620.
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fluorescence intensity versus different p A incubation time. N) Quantification of total
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RNA p A incorporation versus different actinomycin D incubation time in HeLa cells
6 A. M. Smith, M. C. Mancini and S. Nie, Nat. Nanotechnol.,
6
using UHPLC-QQQ-MS/MS. O) Representative mass spectroscopic traces of p A when
2
009, 4, 710.
incubated with actinomycin D for different time. The detection channel is specific to
p6A with mass transition 306 to 174. P) A linear correlation of fluorescence intensity
7 A. T. Aron, K. M. Ramos-Torres, J. A. Cotruvo and C. J. Chang,
Acc. Chem. Res., 2015, 48, 2434.
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changes with p A/A values was derived.
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,
1
y = 149.18x - 11.93
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In summary, we report an assay to visualize and quantify
cellular RNA production and degradation. This approach
provides biological community with a simple, low cost, and
2 X. Gao, J. Z. Sun and B. Z. Tang, Isr. J. Chem., 2018, 58, 1.
3 F. Hu, S. Xu and B. Liu, Adv. Mater., 2018, 30, 1801350.
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robust tool to timely investigate cellular RNA dynamics. 25 F. Hu, X. Cai, P. N. Manghnani, W. Wu and B. Liu, Chem. Sci.,
Compared with commercial fluorescent probes that are prone
2018,
6 Y. Zhao, C. Y. Y. Yu, R. T. K. Kwok, Y. Chen, S. Chen, J. W. Y.
Lam and B. Z. Tang, J. Mater. Chem. B, 2015, , 4993.
9, 2756.
2
2
to photo-bleaching and ACQ, our synthesized SO -TPE-N
3
3
3
probe has AIE property and exhibits high brightness in
biological aqueous environment, outstanding photostability,
and large Stokes shift. During cellular RNA synthesis and decay
7 X. Shu, Q. Dai, T. Wu, I. R. Bothwell, Y. Yue, Z. Zhang, J. Cao,
Q. Fei, M. Luo, C. He and J. Liu, J. Am. Chem. Soc., 2017, 139
17213.
,
processes, the labeled RNA fluorescence signal reveals a linear 28 R. Ottria, S. Casati, E. Baldoli, J. A. M. Maier and P. Ciuffreda,
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Bioorg. Med. Chem., 2010, 18, 8396.
correlation with the change of cellular p A-labeled RNA level
which is validated and quantified by high sensitive mass
spectrometry. Together, our method enables cellular RNA
visualization and quantification in an easy and reliable way.
We thank the National Natural Science Foundation of China
(91853110), National Key Research and Development Program
of China (2017YFA0506800), the Fundamental Research Funds
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| J. Name., 2012, 00, 1-3
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