Skip to main content
SpringerLink
Log in

SYBR Gold Fluorescence Quenching is a Sensitive Probe of Chitosan-microRNA Interactions

  • SHORT COMMUNICATION
  • Published:
Journal of Fluorescence Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

References

  1. Rinaudo M (2006) Chitin and chitosan: properties and applications. Prog Polym Sci 31:603–632. doi:10.1016/j.progpolymsci.2006.06.001

    Article  CAS  Google Scholar 

  2. Harish Prashanth KV, Tharanathan RN (2007) Chitin/chitosan: modifications and their unlimited application potential—an overview. Trends Food Sci Technol 18:117–131. doi:10.1016/j.tifs.2006.10.022

    Article  Google Scholar 

  3. Liu X, Howard KA, Dong M, et al. (2007) The influence of polymeric properties on chitosan/siRNA nanoparticle formulation and gene silencing. Biomaterials 28:1280–1288. doi:10.1016/j.biomaterials.2006.11.004

    Article  PubMed  CAS  Google Scholar 

  4. Lavertu M, Méthot S, Tran-Khanh N, Buschmann MD (2006) High efficiency gene transfer using chitosan/DNA nanoparticles with specific combinations of molecular weight and degree of deacetylation. Biomaterials 27:4815–4824. doi:10.1016/j.biomaterials.2006.04.029

    Article  PubMed  CAS  Google Scholar 

  5. Ragelle H, Vandermeulen G, Préat V (2013) Chitosan-based siRNA delivery systems. J Control Release 172:207–218. doi:10.1016/j.jconrel.2013.08.005

    Article  PubMed  CAS  Google Scholar 

  6. Koping-Hoggard M, Tubulekas I, Guan H, et al. (2001) Chitosan as a nonviral gene delivery system. Structure-property relationships and characteristics compared with polyethylenimine in vitro and after lung administration in vivo. Gene Ther 8:1108–1121. doi:10.1038/sj.gt.3301492

    Article  PubMed  CAS  Google Scholar 

  7. McKiernan PJ, Cunningham O, Greene CM, Cryan S-A (2013) Targeting miRNA-based medicines to cystic fibrosis airway epithelial cells using nanotechnology. Int J Nanomedicine 8:3907–3915. doi:10.2147/IJN.S47551

    PubMed  PubMed Central  Google Scholar 

  8. Santos-Carballal B, Aaldering LJ, Ritzefeld M, et al. (2015) Physicochemical and biological characterization of chitosan-microRNA nanocomplexes for gene delivery to MCF-7 breast cancer cells. Sci Rep 5:13567. doi:10.1038/srep13567

    Article  PubMed  CAS  PubMed Central  Google Scholar 

  9. Adammek M, Greve B, Kässens N, et al. (2013) MicroRNA miR-145 inhibits proliferation, invasiveness, and stem cell phenotype of an in vitro endometriosis model by targeting multiple cytoskeletal elements and pluripotency factors. Fertil Steril 99:1346–1355e5. doi:10.1016/j.fertnstert.2012.11.055

    Article  PubMed  CAS  Google Scholar 

  10. Götte M, Mohr C, Koo C-Y, et al. (2010) MiR-145-dependent targeting of Junctional Adhesion Molecule A and modulation of fascin expression are associated with reduced breast cancer cell motility and invasiveness. Oncogene 29:6569–6580. doi:10.1038/onc.2010.386

    Article  PubMed  Google Scholar 

  11. Liu W, Sun S, Cao Z, et al. (2005) An investigation on the physicochemical properties of chitosan/DNA polyelectrolyte complexes. Biomaterials 26:2705–2711. doi:10.1016/j.biomaterials.2004.07.038

    Article  PubMed  CAS  Google Scholar 

  12. Ma PL, Lavertu M, Winnik FM, Buschmann MD (2009) New insights into chitosan-DNA interactions using isothermal titration microcalorimetry. Biomacromolecules 10:1490–1499. doi:10.1021/bm900097s

    Article  PubMed  CAS  Google Scholar 

  13. Holzerny P, Ajdini B, Heusermann W, et al. (2012) Biophysical properties of chitosan/siRNA polyplexes: profiling the polymer/siRNA interactions and bioactivity. J Control Release 157:297–304. doi:10.1016/j.jconrel.2011.08.023

    Article  PubMed  CAS  Google Scholar 

  14. Wiethoff CM, Gill ML, Koe GS, et al. (2003) A fluorescence study of the structure and accessibility of plasmid DNA condensed with cationic gene delivery vehicles. J Pharm Sci 92:1272–1285. doi:10.1002/jps.10391

    Article  PubMed  CAS  Google Scholar 

  15. Magde D, Elson E, Webb WW (1972) Thermodynamic fluctuations in a reacting system—measurement by fluorescence correlation spectroscopy. Phys Rev Lett 29:705–708. doi:10.1103/PhysRevLett.29.705

    Article  CAS  Google Scholar 

  16. Magde D, Elson EL, Webb WW (1974) Fluorescence correlation spectroscopy. II. An experimental realization. Biopolymers 13:29–61. doi:10.1002/bip.1974.360130103

    Article  PubMed  CAS  Google Scholar 

  17. Mastiholi BM, Tangod VB, Raikar US (2013) Influence of metal nanoparticles on ADS560EI fluorescent laser dye. Opt Int J Light Electron Opt 124:261–264. doi:10.1016/j.ijleo.2011.11.054

    Article  CAS  Google Scholar 

  18. Tangod VB, Raikar P, Mastiholi BM, Raikar US (2014) Solvent polarity studies of highly fluorescent laser dye ADS740WS and its fluorescence quenching with silver nanoparticles. Can J Phys 92:116–123. doi:10.1139/cjp-2013-0250

    Article  CAS  Google Scholar 

  19. Tuma RS, Beaudet MP, Jin X, et al. (1999) Characterization of SYBR gold nucleic acid gel stain: a dye optimized for use with 300-nm ultraviolet transilluminators. Anal Biochem 268:278–288. doi:10.1006/abio.1998.3067

    Article  PubMed  CAS  Google Scholar 

  20. Lakowicz JR (2006) Principles of fluorescence spectroscopy. Springer US, United States

    Book  Google Scholar 

  21. Yu X, Liu R, Yi R, et al. (2011) Study of the interaction between N-confused porphyrin and bovine serum albumin by fluorescence spectroscopy. Spectrochim Acta A Mol Biomol Spectrosc 78:1329–1335. doi:10.1016/j.saa.2011.01.024

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

We acknowledge financial support and a PhD fellowship to BSC from German Research Council DFG (Project GRK 1549 International Research Training Group ‘Molecular and Cellular Glyco-Sciences’); from The Danish Agency for Science, Technology and Innovation, Denmark (FENAMI project 10-093456); the research leading to these results has also received funding from the European Union’s Seventh Framework Program for research, technological development and demonstration under grant agreement n° 613931. We are indebted to Prof. Susanne Fetzner (University of Münster), for her generous access to spectrophotometer and spectrofluorometer instruments. We thank Dr. Juan Pablo Fuenzalida and Dr. Kishore Babu Bobbili for their support during the experiments.

Author information

Authors and Affiliations

  1. Institute of Plant Biology and Biotechnology (IBBP), University of Münster, Schlossgarten 3, D-48149, Münster, Germany

    Beatriz Santos-Carballal, Bruno M. Moerschbacher & Francisco M. Goycoolea

  2. School of Chemistry, University of Hyderabad, Hyderabad, 500046, India

    Musti J. Swamy

Authors
  1. Beatriz Santos-Carballal
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Musti J. Swamy
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bruno M. Moerschbacher
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Francisco M. Goycoolea
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Francisco M. Goycoolea.

Electronic supplementary material

ESM 1

(PDF 729 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Santos-Carballal, B., Swamy, M.J., Moerschbacher, B.M. et al. SYBR Gold Fluorescence Quenching is a Sensitive Probe of Chitosan-microRNA Interactions. J Fluoresc 26, 37–42 (2016). https://doi.org/10.1007/s10895-015-1697-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10895-015-1697-8

Keywords

Search

Navigation