Highly sensitive and selective RNase A recognition systems based on “OFF – ON – OFF” fluorescence probes

The specific recognition and highly sensitive assay of RNase A was conducted based on H1-RNA or L1-RNA fluorescence “Switching-ON” complex, and the detection limit is as low as 2.87 × 10−7 U/mL. [Display omitted] •New fluorescence probes assay for monitoring endoribonuclease activity.•The fluorescen...

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Bibliographic Details
Published in:Sensors and actuators. B, Chemical Chemical, 2018-04, Vol.259, p.282-288
Main Authors: Du, Jinya, Yang, Huiran, Huang, Na, Dong, Yuzhi, Gao, Qingyun, Yang, Wei, Liu, Biao, Yang, Changying
Format: Article
Language:English
Subjects:
Biotechnology
Cloning
Deoxyribonucleic acid
DNA
DNA repair
Endonuclease
Enzyme activity
Fluorescence
Fluorescence probes
Fluorescent indicators
Inhibitor
Recognition
Ribonucleases
Ribonucleic acid
RNA
RNase A
Switching
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Summary:The specific recognition and highly sensitive assay of RNase A was conducted based on H1-RNA or L1-RNA fluorescence “Switching-ON” complex, and the detection limit is as low as 2.87 × 10−7 U/mL. [Display omitted] •New fluorescence probes assay for monitoring endoribonuclease activity.•The fluorescence “Switching – On” complex H1- RNA or L1-RNA exhibited highly specificity and selectivity for the target RNase A.•This method showed high sensitivity with a detection limit of 2.87 × 10−7 U/mL for endoribonuclease RNase A, which is much lower than many other reported.•The enzyme assay is simply done for rapid qualitative and quantitative indication of endoribonuclease activities and inhibition capabilities in aqueous physiology, without separation or isotopic substrates labeling. The “Switching – On” complexes, indolium derivatives bound with nuclear acids, were used as fluorescence probes to recognize the endonuclease and monitor the enzyme activities. Three probes, H1, L1 and N1, weakly emitted in aqueous buffer, exhibited strong red emissions when they bound into DNA or RNA. For H1-RNA or L1-RNA, the fluorescence was sensitively quenched by endoribonuclease, RNase A. The quenching of H1 or L1 originated only from the cleavage of RNase A to RNA, which was proved by various control and interference experiments. The specific recognition and highly sensitive enzyme activity assay of RNase A was simply conducted based on H1-RNA or L1-RNA “Switching-ON” fluorescent complex, and the detection limit (DL) is as low as 2.87 × 10−7 U/mL for H1-RNA, and 5.02 × 10−7 U/mL for L1-RNA, respectively. This approach was also used to evaluate the efficiency of inhibitors for RNase A.
ISSN:0925-4005
1873-3077
DOI:10.1016/j.snb.2017.12.072