Synthesis and label free characterization of a bimolecular PNA homo quadruplex

Biochim Biophys Acta Gen Subj. 2017 May;1861(5 Pt B):1222-1228. doi: 10.1016/j.bbagen.2016.11.037. Epub 2016 Nov 29.

Abstract

Background: G-quadruplex DNA is involved in many physiological and pathological processes. Both clinical and experimental studies on DNA G-quadruplexes are slowed down by their enzymatic instability. In this frame, more stable chemically modified analogs are needed.

Methods: The bis-end-linked-(gggt)2 PNA molecule (BEL-PNA) was synthesized using in solution and solid phase synthetic approaches. Quadruplex formation was assessed by circular dichroism (CD) and surface enhanced Raman scattering (SERS).

Results: An unprecedented bimolecular PNA homo quadruplex is here reported. To achieve this goal, we developed a bifunctional linker that once functionalized with gggt PNA strands and annealed in K+ buffer allowed the obtainment of a PNA homo quadruplex. The identification of the strong SERS band at ~1481cm-1, attributable to vibrations involving the quadruplex diagnostic Hoogsteen type hydrogen bonds, confirmed the formation of the PNA homo quadruplex.

Conclusions: By tethering two G-rich PNA strands to the two ends of a suitable bifunctional linker it is possible to obtain bimolecular PNA homo quadruplexes after annealing in K+-containing buffers. The formation of such CD-unfriendly complexes can be monitored, even at low concentrations, by using the SERS technique.

General significance: Given the importance of DNA G-quadruplexes in medicine and nanotechnology, the obtainment of G-quadruplex analogs provided with enhanced enzymatic stability, and their monitoring by highly sensitive label-free techniques are of the highest importance. This article is part of a Special Issue entitled "G-quadruplex" Guest Editor: Dr. Concetta Giancola and Dr. Daniela Montesarchio.

Keywords: G-quadruplex; Label free monitoring; PNA; SERS; Surface enhanced Raman scattering.

MeSH terms

  • Base Sequence
  • Circular Dichroism
  • G-Quadruplexes*
  • Guanine / chemistry*
  • Hydrogen Bonding
  • Models, Molecular
  • Peptide Nucleic Acids / chemistry*
  • Spectrum Analysis, Raman
  • Structure-Activity Relationship

Substances

  • Peptide Nucleic Acids
  • Guanine