SYBR Gold Fluorescence Quenching is a Sensitive Probe of Chitosan-microRNA Interactions.
Competitive dye displacement titration has previously been used to characterize chitosan-DNA interactions using ethidium bromide. In this work, we aim to develop a fast and reliable method using SYBR Gold as a fluorescent probe to evaluate the binding affinity between ssRNA and chitosan. The interaction of chitosan with ssRNA was investigated as a function of temperature, molecular weight and degree of acetylation of chitosan, using competitive dye displacement titrations with fluorescence quenching. Affinity constants are reported, showing the high sensitivity of the interaction to the degree of acetylation of chitosan and barely dependent on the molecular weight. We propose that the mechanism of SYBR Gold fluorescence quenching is governed by both static and dynamic quenching.
Institute of Plant Biology and Biotechnology (IBBP), University of Münster, Schlossgarten 3, D-48149, Münster, Germany.
Beatriz
B
Santos-Carballal
School of Chemistry, University of Hyderabad, Hyderabad, 500046, India.
Musti J
MJ
Swamy
Institute of Plant Biology and Biotechnology (IBBP), University of Münster, Schlossgarten 3, D-48149, Münster, Germany.
Bruno M
BM
Moerschbacher
Institute of Plant Biology and Biotechnology (IBBP), University of Münster, Schlossgarten 3, D-48149, Münster, Germany. goycoole@uni-muenster.de.
Francisco M
FM
Goycoolea
D048271:Chitosan / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D035683:MicroRNAs / Q000737:chemistry*; D009930:Organic Chemicals / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D005456:Fluorescent Dyes; D035683:MicroRNAs; D009930:Organic Chemicals; C474470:SYBR Gold nucleic acid gel stain; D048271:Chitosan
Chitosan; Dye displacement titration; Fluorescence quenching; Single-stranded RNA