title: Multicolor fluorescent biosensor for multiplexed detection of DNA.
abstract: Development of efficient methods for highly sensitive and rapid screening of specific oligonucleotide sequences is essential to the early diagnosis of serious diseases. In this work, an aggregated cationic perylene diimide (PDI) derivative was found to efficiently quench the fluorescence emission of a variety of anionic oligonucleotide-labeled fluorophores that emit at wavelengths from the visible to NIR region. This broad-spectrum quencher was then adopted to develop a multicolor biosensor via a label-free approach for multiplexed fluorescent detection of DNA. The aggregated perylene derivative exhibits a very high quenching efficiency on all ssDNA-labeled dyes associated with biosensor detection, having efficiency values of 98.3 ± 0.9%, 97 ± 1.1%, and 98.2 ± 0.6% for FAM, TAMRA, and Cy5, respectively. An exonuclease-assisted autocatalytic target recycling amplification was also integrated into the sensing system. High quenching efficiency combined with autocatalytic target recycling amplification afforded the biosensor with high sensitivity toward target DNA, resulting in a detection limit of 20 pM, which is about 50-fold lower than that of traditional unamplified homogeneous fluorescent assay methods. The quencher did not interfere with the catalytic activity of nuclease, and the biosensor could be manipulated in either preaddition or postaddition manner with similar sensitivity. Moreover, the proposed sensing system allows for simultaneous and multicolor analysis of several oligonucleotides in homogeneous solution, demonstrating its potential application in the rapid screening of multiple biotargets.
authors.affiliation: Molecular Science and Biomedicine Laboratory, State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, Collaborative Innovation Center of Molecular Engineering for Theranostics, Hunan University , Changsha 410082, P.R. China.
authors.forename: Rong
authors.initials: R
authors.lastname: Hu
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.forename: Xiao-Bing
authors.initials: XB
authors.lastname: Zhang
authors.forename: Shuang-Yan
authors.initials: SY
authors.lastname: Huan
authors.forename: Cuichen
authors.initials: C
authors.lastname: Wu
authors.forename: Ting
authors.initials: T
authors.lastname: Fu
authors.forename: Weihong
authors.initials: W
authors.lastname: Tan
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation*; D004247:DNA / Q000737:chemistry*; D004277:DNA, Single-Stranded / Q000737:chemistry; D004720:Endonucleases / Q000737:chemistry; D005456:Fluorescent Dyes; D059014:High-Throughput Nucleotide Sequencing; D009841:Oligonucleotides / Q000737:chemistry
chemical_list: D004277:DNA, Single-Stranded; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D004247:DNA; D004720:Endonucleases
references.citation: Anal Chem. 2012 Dec 18;84(24):10845-51
references.pmid: 23167838
references.citation: Anal Chem. 2013 Feb 19;85(4):2282-8
references.pmid: 23320625
references.citation: Science. 1997 Aug 22;277(5329):1078-81
references.pmid: 9262471
references.citation: J Biol Chem. 1964 Jan;239:251-8
references.pmid: 14114851
references.citation: Chem Soc Rev. 2010 Oct;39(10):3746-71
references.pmid: 20820469
references.citation: Anal Chem. 2011 Jan 1;83(1):14-7
references.pmid: 21117628
references.citation: Bioorg Med Chem Lett. 2002 Dec 2;12(23):3395-8
references.pmid: 12419369
references.citation: Nucleic Acids Res. 2004 Jul 19;32(13):e108
references.pmid: 15263064
references.citation: J Am Chem Soc. 2001 Jul 11;123(27):6726-7
references.pmid: 11439073
references.citation: Chem Soc Rev. 2010 Nov;39(11):4234-43
references.pmid: 20871878
references.citation: Nat Biotechnol. 2001 Apr;19(4):365-70
references.pmid: 11283596
references.citation: Angew Chem Int Ed Engl. 2013 Mar 11;52(11):3177-80
references.pmid: 23386400
references.citation: Anal Chem. 2012 Jun 19;84(12):5216-20
references.pmid: 22721426
references.citation: PLoS One. 2012;7(2):e31126
references.pmid: 22363564
references.citation: J Am Chem Soc. 2008 May 28;130(21):6820-5
references.pmid: 18459781
references.citation: Anal Chem. 2008 May 15;80(10):3832-7
references.pmid: 18407673
references.citation: Nat Biotechnol. 1998 Jan;16(1):49-53
references.pmid: 9447593
references.citation: Angew Chem Int Ed Engl. 2010 Feb 15;49(8):1485-8
references.pmid: 20094998
references.citation: Science. 1996 Sep 13;273(5281):1516-7
references.pmid: 8801636
references.citation: Anal Chem. 2009 Jan 15;81(2):656-61
references.pmid: 19093760
references.citation: Anal Chem. 2012 Apr 17;84(8):3568-73
references.pmid: 22420689
references.citation: J Am Chem Soc. 2011 Nov 9;133(44):17524-7
references.pmid: 21954932
references.citation: J Am Chem Soc. 2007 Oct 3;129(39):11896-7
references.pmid: 17850085
references.citation: Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12287-92
references.pmid: 10535914
references.citation: Chem Rev. 2009 May;109(5):1948-98
references.pmid: 19301873
references.citation: Angew Chem Int Ed Engl. 2012 Feb 13;51(7):1625-30
references.pmid: 22213412
references.citation: Nanoscale. 2013 Jan 21;5(2):619-23
references.pmid: 23208411
references.citation: Anal Chem. 2011 Jul 1;83(13):5062-6
references.pmid: 21639104
references.citation: Chem Rev. 2008 Feb;108(2):423-61
references.pmid: 18229952
references.citation: Chem Rev. 2003 Nov;103(11):4419-76
references.pmid: 14611267
references.citation: J Am Chem Soc. 2008 Jul 2;130(26):8351-8
references.pmid: 18528999
references.citation: J Am Chem Soc. 2007 Aug 29;129(34):10394-401
references.pmid: 17676734