分子信标的构建及其应用研究进展

doi:10.3724/SP.J.1095.2013.30116

摘要/Abstract

摘要： 分子信标（molecular beacon,MB）是一种寡聚核苷酸荧光探针，其具有灵敏度高、特异性强、操作简单以及不必与未反应的探针分离即可实时检测等优点，在分子生物学和基因组学及分子医学等领域具有十分重要的应用价值。本文介绍了近年来出现的各种新型分子信标的结构及工作原理，概述了分子信标技术在生命科学领域中的应用，展望了分子信标技术的发展趋势。

关键词: 分子信标, 构建, 生化分析, 应用

Abstract: Molecular beacon is an oligonucleotide probe of fluorescence. Molecular beacons are playing a very important role in molecular biology, genetics and molecular medicine because they have been shown to have very high selectivity, high sensitivity, and real-time detection capabilities. The structures and working mechanisms of a variety of novel molecular beacons have been outlined here. We briefly summarize the application of molecular beacon technology in the field of life sciences, and propose the prospect this technology.

Key words: molecular beacons, construction, biochemical analysis, application

中图分类号:

O651

向东山, 翟琨. 分子信标的构建及其应用研究进展[J]. 应用化学, DOI: 10.3724/SP.J.1095.2013.30116.

XIANG Dongshan1,2*, ZHAI Kun2. Construction of Molecular Beacons and Their Research Progress of Application[J]. Chinese Journal of Applied Chemistry, DOI: 10.3724/SP.J.1095.2013.30116.

参考文献

[1] Tyagi S,Kramer F R. Molecular Beacons:Probes that Fluoresce upon Hybridization[J]. Nat Biotech,1996,14(3):303-308.

[2] Lv C,Yu L L,Wang J,et al. A Dumbbell Molecular Beacon for the Specific Recognition of Nucleic Acids[J]. Bioorg Med Chem Lett,2010,20(22):6547-6550.

[3] Haner R,Biner S M,Langenegger S M,et al. A Highly Sensitive, Excimer-controlled Molecular Beacon[J]. Angew Chem Int Ed,2010,49(7):1227-1230.

[4] Peng X H,Cao Z H,Xia J T,et al. Real-time Detection of Gene Expression in Cancer Cells Using Molecular Beacon Imaging:New Strategies for Cancer Research[J]. Cancer Res,2005,65(5):1909-1917.

[5] Li N,Wong P K. Transfection of Molecular Beacons in Microchannels for Single-cell Gene-expression Analysis[J]. Bioanalysis,2010,2(10):1689-1699.

[6] Singh J,Batish V K,Grover S. Molecular Beacon Based Real-time PCR Assay for Simultaneous Detection of Listeria Monocytogenes and Salmonella Spp in Dairy Products[J]. Dairy Sci Technol,2011,91(3):373-382.

[7] Li Y Q,Guan L Y,Wang J H,et al. Simultaneous Detection of Dual Single-base Mutations by Capillary Electrophoresis Using Quantum Dot-molecular Beacon Probe[J]. Biosens Bioelectron,2011,26(5):2317-2322.

[8] Wu C S,Oo M K K,Cupps J M,et al. Robust Silica-coated Quantum Dot-molecular Beacon for Highly Sensitive DNA Detection[J]. Biosens Bioelectron,2011,26(9):3870-3875.

[9] Kong R M,Zhang X B,Zhang L L,et al. Molecular Beacon-based Junction Probes for Efficient Detection of Nucleic Acids via a True Target-triggered Enzymatic Recycling Amplification[J]. Anal Chem,2011,83(1):14-17.

[10] Nitin N,Santangelo P J,Kim G,et al. Peptide-linked Molecular Beacons for Efficient Delivery and Rapid mRNA Detection in Living Cells[J]. Nucl Acids Res,2004,32(6):e58.

[11] Liu B,Yang X H,Wang K M,et al. Quantitative Detection of p21 mRNA in Tumor Cells Based on Molecular Beacon[J]. Acta Chim Sin,2010,68(13):1303-1307.

[12] Tang D,Liao D L,Zhu Q K,et al. Fluorescence Turn-on Detection of a Protein Through the Displaced Single-stranded DNA Binding Protein Binding to a Molecular Beacon[J]. Chem Commun,2011,47(19):5485-5487.

[13] Ganguli P S,Chen W,Yates M V. Detection of Murine Norovirus-1 by Using TAT Peptide-delivered Molecular Beacons[J]. Appl Environ Microbiol,2011,77(15):5517-5520.

[14] Bercovici M,Kaigala G V,Mach K E,et al. Rapid Detection of Urinary Tract Infections Using Isotachophoresis and Molecular Beacons[J]. Anal Chem,2011,83(11):4110-4117.

[15] Peng Z Y,Soper S A,Pingle M R,et al. Ligase Detection Reaction Generation of Reverse Molecular Beacons for Near Real-time Analysis of Bacterial Pathogens Using Single-pair Fluorescence Resonance Energy Transfer and a Cyclic Olefin Copolymer Microfluidic Chip[J]. Anal Chem,2010,82(23):9727-9735.

[16] Yeh H Y,Hwang Y C,Yates M V,et al. Detection of Hepatitis A Virus by Using a Combined Cell Culture-molecular Beacon Assay[J]. Appl Environ Microbiol,2008,74(7):2239-2243.

[17] Ma C B,Tang Z W,Huo X Q,et al. Real-time Monitoring of Double-stranded DNA Cleavage Using Molecular Beacons[J]. Talanta,2008,76(2):458-461.

[18] Meng X X,Wang K M,Tan W H,et al. Ribozyme Probe Based on Molecular Beacon for Real Time Monitoring of Enzymatic Cleavage Process[J]. Chinese Sci Bull,2003,48(23):2581-2584.

[19] Meng X X,Tang Z W,Wang K M,et al. Ultrasensitive Monitoring of Ribozyme Cleavage Product Using Molecular-beacon-ligation System[J]. Chinese Sci Bull,2007,52(5):603-607.

[20] Wang J,Onoshima D,Aki M,et al. Label-free Detection of DNA-binding Proteins Based on Microfluidic Solid-state Molecular Beacon Sensor[J]. Anal Chem,2011,83(9):3528-3532.

[21] Hara Y,Fujii T,Kashida H,et al. Coherent Quenching of a Fluorophore for the Design of a Highly Sensitive In-stem Molecular Beacon[J]. Angew Chem Int Ed,2010,49(32):5502-5506.

[22] Wang K M,Tang Z W,Yang C Y J,et al. Molecular Engineering of DNA:Molecular Beacons[J]. Angew Chem Int Ed,2009,48(5):856-870.

[23] Gifford L K,Jordan D,Pattanayak V,et al. Stemless Self-quenching Reporter Molecules Identify Target Sequences in mRNA[J]. Anal Biochem,2005,347(1):77-88.

[24] Maxwell D J,Taylor J R,Nie S M. Self-assembled Nanoparticle Probes for Recognition and Detection of Biomolecules[J]. J Am Chem Soc,2002,124(32):9606-9612.

[25] Dubertret B,Calame M,Libchaber A J. Single-mismatch Detection Using Gold-quenched Fluorescent Oligonucleotides[J]. Nat Biotechnol,2001,19(7):680-681.

[26] Song S P,Liang Z Q,Zhang J,et al. Gold-nanoparticle-based Multicolor Nanobeacons for Sequence-specific DNA Analysis[J]. Angew Chem Int Ed,2009,48(46):8670-8674.

[27] Brunner J,Kraemer R. Copper(II)-quenched Oligonucleotide Probes for Fluorescent DNA Sensing[J]. J Am Chem Soc,2004,126(42):13626-13627.

[28] Yang C Y J,Lin H,Tan W H. Molecular Assembly of Superquenchers in Signaling Molecular Interactions[J]. J Am Chem Soc,2005,127(37):12772-12773.

[29] Kurata S,Kanagawa T,Yamada K,et al. Fluorescent Quenching-based Quantitative Detection of Specific DNA/RNA Using a BODIPY(R) FL-labeled Probe or Primer[J]. Nucl Acids Res,2001,29(6):e34.

[30] Lu C H,Li J,Liu J J,et al. Increasing the Sensitivity and Single-base Mismatch Selectivity of the Molecular Beacon Using Graphene Oxide as the “Nanoquencher”[J]. Chem-Eur J,2010,16(16):4889-4894.

[31] Xiang D S,Zheng A H,Luo M,et al. Graphene Oxide and Molecular Beacons-based Multiplexed DNA Detection by Synchronous Fluorescence Analysis[J]. Sci China Chem,2013,56(3):380-386.

[32] Conlon P,Yang C Y J,Wu Y R,et al. Pyrene Excimer Signaling Molecular Beacons for Probing Nucleic Acids[J]. J Am Chem Soc,2008,130(1):336-342.

[33] Chi C W,Lao Y H,Li Y S,et al. A Quantum Dot-aptamer Beacon Using a DNA Intercalating Dye as the FRET Reporter:Application to Label-free Thrombin Detection[J]. Biosens Bioelectron,2011,26(7):3346-3352.

[34] Zhang P,Beck T,Tan W H. Design of a Molecular Beacon DNA Probe with Two Fluorophores[J]. Angew Chem Int Ed,2001,40(2):402-405.

[35] Tyagi S,Marras S A E,Kramer F R. Wavelength-shifting Molecular Beacons[J]. Nat Biotechnol,2000,18(11):1191-1196.

[36] Tsourkas A,Behlke M A,Xu Y Q,et al. Spectroscopic Features of Dual Fluorescence/Luminescence Resonance Energy-transfer Molecular Beacons[J]. Anal Chem,2003,75(15):3697-3703.

[37] Wang Y X,Li J S,Jin J Y,et al. Strategy for Molecular Beacon Binding Readout:Separating Molecular Recognition Element and Signal Reporter[J]. Anal Chem,2009,81(23):9703-9709.

[38] Gifford L K,Jordan D,Pattanayak V,et al. Stemless Self-quenching Reporter Molecules Identify Target Sequences in mRNA[J]. Anal Biochem,2005,347(1):77-88.

[39] Tsourkas A,Behlke M A,Bao G. Structure-function Relationships of Shared-stem and Conventional Molecular Beacons[J]. Nucl Acids Res,2002,30(19):4208-4215.

[40] Wang L,Yang C Y J,Medley C D,et al. Locked Nucleic Acid Molecular Beacons[J]. J Am Chem Soc,2005,127(45):15664-15665.

[41] Wang W J,Chen C L,Qian M X,et al. Aptamer Biosensor for Protein Detection Based on Guanine-quenching[J]. Sens Actuators B,2008,129(1):211-217.

[42] Li J W J,Fang X H,Tan W H. Molecular Aptamer Beacons for Real-time Protein Recognition[J]. Biochem Biophys Res Commun,2002,292(1):31-40.

[43] Tan X H,Chen W J,Lu S,et al. Molecular Beacon Aptamers for Direct and Universal Quantitation of Recombinant Proteins from Cell Lysates[J]. Anal Chem,2012,84(19):8272-8276.

[44] Zhuang H L,Zhen S J,Wang J,et al. Sensitive Detection of Prion Protein Through Long Range Resonance Energy Transfer Between Graphene Oxide and Molecular Aptamer Beacon[J]. Anal Method,2013,5(1):208-212.

[45] Guetschow E D,Black W,Walsh C M,et al. Detection of Prolactin Inducible Protein mRNA, a Biomarker for Breast Cancer Metastasis, Using a Molecular Beacon-based Assay[J]. Anal Bioanal Chem,2012,404(2):399-406.

[46] Jiang Y L,McGoldrick C A,Yin D L,et al. A Specific Molecular Beacon Probe for the Detection of Human Prostate Cancer Cells[J]. Bioorg Med Chem Lett,2012,22(11):3632-3638.

[47] Shukoor M I,Altman M O,Han D,et al. Aptamer-nanoparticle Assembly for Logic-based Detection[J]. Acs Appl Mater Inter,2012,4(6):3007-3011.

[48] Lin Y W,Chang H T. Detection of Mercury and Phenylmercury Ions Using DNA-based Fluorescent Probe[J]. Analyst,2011,136(16):3323-3328.

[49] Gao Y,Li Y,Zou X,et al. Highly Sensitive and Selective Detection of Biothiols Using Graphene Oxide-based “Molecular Beacon”-Like Fluorescent Probe[J]. Anal Chim Acta,2012,731:68-74.

[50] Qi L,Zhao Y,Yuan H,et al. Amplified Fluorescence Detection of Mercury(Ⅱ) Ions (Hg2+) Using Target-induced DNAzyme Cascade with Catalytic and Molecular Beacons[J]. Analyst,2012,137(12):2799-2805.

[51] Stobiecka M,Molinero A A,Chalupa A,et al. Mercury/Homocysteine Ligation-induced ON/OFF-Switching of a T-T Mismatch-based Oligonucleotide Molecular Beacon[J]. Anal Chem,2012,84(11):4970-4978.

[52] Zhang M,Yin B C,Tan W,et al. A Versatile Graphene-based Fluorescence “On/Off” Switch for Multiplex Detection of Various Targets[J]. Biosens Bioelectron,2011,26(7):3260-3265.

[53] Biggins J B,Prudent J R,Marshall D J,et al. A Continuous Assay for DNA Cleavage:The Application of “Break Lights” to Enediynes, Iron-dependent Agents, and Nucleases[J]. Proc Natl Acad Sci USA,2000,97(25):13537-13542.

[54] He Y,Wang Z G,Tang H W,et al. Low Background Signal Platform for the Detection of ATP:When a Molecular Aptamer Beacon Meets Graphene Oxide[J]. Biosens Bioelectron,2011,29(1):76-81.

[55] Ke G L,Wang C,Ge Y,et al. L-DNA Molecular Beacon:A Safe, Stable, and Accurate Intracellular Nano-thermometer for Temperature Sensing in Living Cells[J]. J Am Chem Soc,2012,134(46):18908-18911.