title: A Nonenzymatic Hairpin DNA Cascade Reaction Provides High Signal Gain of mRNA Imaging inside Live Cells.
abstract: Enzyme-free signal amplification has enabled sensitive in vitro detection of biomolecules such as proteins and nucleic acids. However, monitoring targets of interest in live cells via enzyme-free amplification is still challenging, especially for analytes with low concentrations. To the best of our knowledge, this paper reports the first attempt to perform mRNA imaging inside live cells, using a nonenzymatic hairpin DNA cascade reaction for high signal gain, termed a hairpin DNA cascade amplifier (HDCA). In conventional nucleic acid probes, such as linear hybridization probes, mRNA target signaling occurs in an equivalent reaction ratio (1:1), whereas, in HDCA, one mRNA target is able to yield multiple signal outputs (1:m), thus achieving the goal of signal amplification for low-expression mRNA targets. Moreover, the recycled mRNA target in the HDCA serves as a catalyst for the assembly of multiple DNA duplexes, generating the fluorescent signal of reduced MnSOD mRNA expression, thus indicating amplified intracellular imaging. This programmable cascade reaction presents a simple and modular amplification mechanism for intracellular biomarkers of interest, providing a significant boost to the search for clues leading to the accurate identification and effective treatment of cancers.
authors.affiliation: ‡Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.|†Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Cuichen
authors.initials: C
authors.lastname: Wu
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Sena
authors.initials: S
authors.lastname: Cansiz
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Liqin
authors.initials: L
authors.lastname: Zhang
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: I-Ting
authors.initials: IT
authors.lastname: Teng
authors.affiliation: ‡Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Qiu
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Juan
authors.initials: J
authors.lastname: Li
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Cuisong
authors.initials: C
authors.lastname: Zhou
authors.affiliation: ‡Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.
authors.forename: Rong
authors.initials: R
authors.lastname: Hu
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Tao
authors.initials: T
authors.lastname: Zhang
authors.affiliation: †Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Cheng
authors.initials: C
authors.lastname: Cui
authors.affiliation: ‡Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Cui
authors.affiliation: ‡Molecular Science and Biomedicine Laboratory, State Key Laboratory for Chemo/Bio Sensing and Chemometrics, College of Chemistry and Chemical Engineering, College of Biology, and Collaborative Research Center of Molecular Engineering for Theranostics, Hunan University, Changsha 410082, China.|†Center for Research at Bio/Nano Interface, Department of Chemistry and Department of Physiology and Functional Genomics, Health Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, Florida 32611-7200, United States.
authors.forename: Weihong
authors.initials: W
authors.lastname: Tan
mesh_terms: D045744:Cell Line, Tumor; D004247:DNA / Q000737:chemistry*; D006801:Humans; D021141:Nucleic Acid Amplification Techniques; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis* / Q000737:chemistry*
chemical_list: D012333:RNA, Messenger; D004247:DNA
references.citation: Proc Natl Acad Sci U S A. 2013 Apr 2;110(14):5386-91
references.pmid: 23509255
references.citation: Science. 2007 Nov 16;318(5853):1121-5
references.pmid: 18006742
references.citation: Nucleic Acids Res. 2004;32(6):e57
references.pmid: 15084672
references.citation: Proc Natl Acad Sci U S A. 2004 Oct 26;101(43):15275-8
references.pmid: 15492210
references.citation: Nucleic Acids Res. 1992 Apr 11;20(7):1691-6
references.pmid: 1579461
references.citation: Proc Natl Acad Sci U S A. 1993 Apr 1;90(7):3113-7
references.pmid: 8464931
references.citation: J Am Chem Soc. 2005 Nov 16;127(45):15664-5
references.pmid: 16277483
references.citation: Nature. 2008 Jan 17;451(7176):318-22
references.pmid: 18202654
references.citation: Angew Chem Int Ed Engl. 2008;47(34):6330-7
references.pmid: 18680110
references.citation: Nat Methods. 2009 May;6(5):331-8
references.pmid: 19404252
references.citation: Nature. 2010 Jun 10;465(7299):736-45
references.pmid: 20535203
references.citation: Nat Biotechnol. 2010 Nov;28(11):1208-12
references.pmid: 21037591
references.citation: Nucleic Acids Res. 2011 Sep 1;39(16):e110
references.pmid: 21693555
references.citation: J Am Chem Soc. 2012 Nov 21;134(46):18908-11
references.pmid: 23126671
references.citation: ACS Nano. 2013 Mar 26;7(3):2312-9
references.pmid: 23397942
references.citation: J Am Chem Soc. 2013 May 22;135(20):7430-3
references.pmid: 23647466
references.citation: J Am Chem Soc. 2013 Jul 17;135(28):10512-8
references.pmid: 23795550
references.citation: ACS Nano. 2013 Jul 23;7(7):5724-31
references.pmid: 23746078
references.citation: J Am Chem Soc. 2013 Sep 4;135(35):12952-5
references.pmid: 23931073
references.citation: Angew Chem Int Ed Engl. 2014 Feb 24;53(9):2389-93
references.pmid: 24469913
references.citation: Chemistry. 2014 May 12;20(20):5866-73
references.pmid: 24692306
references.citation: Acc Chem Res. 2014 Jun 17;47(6):1825-35
references.pmid: 24828239
references.citation: Chem Soc Rev. 2014 May 21;43(10):3324-41
references.pmid: 24643375
references.citation: Nucleic Acids Res. 2000 Jun 15;28(12):E63
references.pmid: 10871386