SEARCHOTHERbiomedical research, research methods

other·biomedical research, research methods, food science, preclinical research·PMC10164568

Cavity hairpin ThT-light nucleic acid switches: the construction of label- and enzyme-free sensing and imaging platforms

Nucleic Acids Research · 9 authors, 15 centres

AI SUMMARY

FIDELITY 100%

This summary was generated by AI from a single paper. It has not been reviewed by a clinician and is not clinical advice. Verify against the source before acting on it.

AGENT 01 · STRUCTURED SUMMARY

TRACE TO SOURCE

“

TL;DR

This study designed CGG-AAA mismatched cavity hairpin nucleic acid switches (CHTLNAS) that excite ThT fluorescence over 500-fold higher than spontaneous levels. CHTLNAS enabled label- and enzyme-free detection of tetracycline and berberine at the pM level within 10 min and facilitated intracellular ThT imaging.

Full summary

1,208 CHARS

The study designed and validated CGG-AAA mismatched cavity hairpin ThT-light nucleic acid switches (CHTLNAS) for ThT fluorescence excitation and sensing applications. Molecular docking simulations predicted that CGG-AAA cavity structures restrict ThT aromatic ring rotation, enabling fluorescence. Experimental validation showed fluorescence over 500-fold higher than ThT spontaneous, 17 to 20-fold higher than ssDNA, and 2.5 to 5-fold higher than complementary duplex. Based on sequence programmability and allosteric flexibility, the authors constructed three label- and enzyme-free fluorescent/ratiometric sensing platforms targeting tetracycline and berberine aptamers, achieving pM-level detection in milk or plasma within 10 min. Binding affinity of CHTLNAS for ThT (Kd approximately 2.938 microM) was substantially higher than complementary duplex (Kd approximately 159.6 microM). A liposome-modified intracellular tracer platform enabled ThT imaging at nM entry concentrations and prolonged nuclear-entry time over 8 h. Limitations include reliance on in vitro validation without animal or human studies, and the potential existence of superior cavity conformations was acknowledged but not explored.

STUDY TYPE

other

SPECIALTY

biomedical research

SUMMARISED BY

AI pipeline

FIDELITY CHECK

100% · A

ORIGINAL PAPER

PMC10164568

Cavity hairpin ThT-light nucleic acid switches: the construction of label- and enzyme-free sensing and imaging platforms

Nucleic Acids Research

READ THE FULL PAPER

ABSTRACT

Abstract Thioflavin T (ThT) is a classical fluorescent dye gaining prominence in current research regarding nucleic acid conformations (NACs). However, most NACs with the ability to excite ThT fluorescent are unique or form in demanding conditions, limiting the extensiveness and depth of ThT application in sensing and imaging. Therefore, this study proposed CGG-AAA mismatched cavity hairpin ThT-light nucleic acid switches (CHTLNAS) with excellent fluorescence excitation over 500-fold higher than spontaneous, 17∼20-fold higher than ssDNA and 2.5∼5-fold higher than complementary duplex. Based on the excellent fluorescence excitation, convenient conformation formation, good sequence programmability, and flexible allosteric ability (known as the Worm-crack pod mechanism mediated by the target), it achieved the label- and enzyme-free detection of tetracycline (TET) and berberine (BB) at the pM level within 10 min. Moreover, it was found enable to realize the sensitive tracking of intracellular carriers at the nM level of ThT entry concentration, and prolongated its cell nuclear-entry time of ThT over 8 h, overcoming the non-specific high background signal interference of ThT in the nuclear region, and expanding the diversified application of ThT in cell biology research. Therefore, CHTLNAS is a more universal, practical tool than G-quadruplex or other kinds of NACs for ThT development and utilization in sensing and imaging platforms.

CITATION EXPORT

Xinyue Lan, Longjiao Zhu, Yangzi Zhang et al.. Cavity hairpin ThT-light nucleic acid switches: the construction of label- and enzyme-free sensing and imaging platforms. Nucleic Acids Research. (2023). https://doi.org/10.1093/nar/gkad179

AUTHORS

9 · 15 CENTRES

Xinyue Lan, Longjiao Zhu, Yangzi Zhang, Keren Chen, Jia Wang, Zaihui Du, Shuting Li, Xu Chen, Wentao Xu

1 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
2 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100083, China
3 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
4 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
5 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100083, China
6 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
7 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100083, China
8 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
9 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
10 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100083, China
11 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
12 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100083, China
13 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China
14 Key Laboratory of Safety Assessment of Genetically Modified Organism (Food Safety), College of Food Science and Nutritional Engineering, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100083, China
15 Food Laboratory of Zhongyuan, Key Laboratory of Precision Nutrition and Food Quality, Department of Nutrition and Health, China Agricultural University, No. 17, Qinghua East Road, Beijing, 100193, China