title: Labeling-free fluorescent detection of DNA hybridization through FRET from pyrene excimer to DNA intercalator SYBR green I.
abstract: A novel labeling-free fluorescence complex probe has been developed for DNA hybridization detection based on fluorescence resonance energy transfer (FRET) mechanism from pyrene excimer of pyrene-functionalized poly [2-(N, N-dimethylamino) ethyl methacrylate] (PFP) to SYBR Green I (SG, a specific intercalator of double-stranded DNA) in a cost-effective, rapid and simple manner. The complex probe consists of the positively charged PFP, SG and negatively charged single-stranded DNA (ssDNA). Upon adding a complementary strand to the complex probe solution, double-stranded DNA (dsDNA) was formed, followed by the intercalation of SG into dsDNA. The pyrene excimer emission was overlapped with the absorption of SG very well and the electrostatic interactions between PFP and dsDNA kept them in close proximity, enabling efficient FRET from pyrene excimer to SG. The fluorescence of SG in the duplex DNA resulting from FRET can be successfully applied to detect DNA hybridization with high sensitivity for a very low detection limit of 10nM and excellent selectivity for detection of single base pair mismatch.
authors.affiliation: School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083, China.
authors.forename: Ruyi
authors.initials: R
authors.lastname: Zhou
authors.affiliation: School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Xu
authors.affiliation: School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Dong
authors.affiliation: School of Materials Science and Engineering, University of Science and Technology Beijing, Beijing 100083, China. Electronic address: guojie.wang@mater.ustb.edu.cn.
authors.forename: Guojie
authors.initials: G
authors.lastname: Wang
mesh_terms: D052160:Benzothiazoles; D004247:DNA / Q000032:analysis*; D003959:Diamines; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D007364:Intercalating Agents / Q000737:chemistry*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D009930:Organic Chemicals / Q000737:chemistry*; D011721:Pyrenes / Q000737:chemistry*; D011804:Quinolines
chemical_list: D052160:Benzothiazoles; D003959:Diamines; D005456:Fluorescent Dyes; D007364:Intercalating Agents; D009930:Organic Chemicals; D011721:Pyrenes; D011804:Quinolines; C098022:SYBR Green I; D004247:DNA; C030984:pyrene
keywords: Biosensor; DNA hybridization; FRET; Fluorescence; Pyrene excimer