Label-Free Probing of G-Quadruplex Formation by Surface-Enhanced Raman Scattering

Rusciano, Giulia; Luca, Anna Chiara De; Pesce, Giuseppe; Sasso, Antonio; Oliviero, Giorgia; Amato, Jussara; Borbone, Nicola; D’Errico, Stefano; Piccialli, Vincenzo; Piccialli, Gennaro; Mayol, Luciano

doi:10.1021/ac201783h

Cited by 54 publications

(60 citation statements)

References 41 publications

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“…Studies have shown that SERS can be used to analyse G-quadruplex and duplex sequences without the use of Raman reporters, however this was performed using a laser excitation frequency of 785 nm and the addition of an aggregating agents such as magnesium sulphate (MgSO 4 ) or spermine. 28,41 Here the SERS measurements were performed using a laser excitation of 532 nm in the absence of any aggregating agent. In the spectrum of 3AQN, the peak at 1380 cm -1 was used for comparison studies, the peak at 719 cm -1 in the 6AQN spectrum was chosen and for 360A peak intensity studies, the unique peak at 1379 cm -1 was used.…”

Section: Resultsmentioning

confidence: 99%

“…Rusciano et al then further developed the application of SERS as a method to determine the stability of G-quadruplexes by using direct SERS analysis to gain information on the structure of guanine-rich DNA sequences. 30 Different G-quadruplex structures were analysed and their relative stabilities were compared. Specific Raman features of G-quadruplex DNAs were identified in the SERS spectra and subsequent analysis on the phosphate backbone conformation and the strength of the Hoogsteen-like hydrogen bonding was performed.…”

Section: Introductionmentioning

confidence: 99%

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Qualitative SERS analysis of G-quadruplex DNAs using selective stabilising ligands

Gracie

Dhamodharan

Pradeepkumar

et al. 2014

Analyst

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Nucleic acids are of key biological importance due to their range of functions and ability to form various different structures, with an example of emerging significance being quadruplexes formed by guanine-rich sequences. These guanine rich sequences are found in different regions of the genome such as telomeres, gene promoters and introns and UTRs of mRNAs. Here a new approach has been developed that utilises surface enhanced Raman scattering (SERS) for the detection of the formation of G-quadruplexes. Three G-quadruplex stabilising ligands that each have their own unique SERS response were used in this study and their ability to act as reporters assessed. A SERS response was only obtained from the ligands in the absence of G-quadruplex formation. This resulted in an "on/off" method which was successfully used to qualitatively detect the formation of G-quadruplex using quadruplex-forming sequences such as human telomeric and C-MYC promoter DNAs. The unique SERS spectra of each stabilising ligand offer the potential for use of SERS to study higher order DNA structures. This work shows that the ligands used can act simultaneously as a potential therapeutic stabilising agent and a SERS reporter, therefore allowing the use of SERS as a method of analysis of the formation of G-quadruplex DNAs.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Qualitative SERS analysis of G-quadruplex DNAs using selective stabilising ligands

Gracie

Dhamodharan

Pradeepkumar

et al. 2014

Analyst

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“…The SERS EF depends on several parameters, such as the real and imaginary parts of the dielectric function of the metal nanostructure, the active substrate's geometry, the metal-molecule affinity, and the possible presence of electronic transition resonances of the analyte molecule at the excitation wavelength. Despite such complexity, SERS has found successful application in cutting-edge biochemical and medical research [20][21][22]. In particular, SERS technique has proved to be an effective platform for optical probing and imaging of live cells via SERS nanotags or in label-free experiments for surface cell sensing [23][24][25][26][27][28][29].…”

Section: Introductionmentioning

confidence: 99%

Cell Imaging by Spontaneous and Amplified Raman Spectroscopies

Rusciano

Zito

Pesce

et al. 2017

Journal of Spectroscopy

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Raman spectroscopy (RS) is a powerful, noninvasive optical technique able to detect vibrational modes of chemical bonds. The high chemical specificity due to its fingerprinting character and the minimal requests for sample preparation have rendered it nowadays very popular in the analysis of biosystems for diagnostic purposes. In this paper, we first discuss the main advantages of spontaneous RS by describing the study of a single protozoan (Acanthamoeba), which plays an important role in a severe ophthalmological disease (Acanthamoeba keratitis). Later on, we point out that the weak signals that originated from Raman scattering do not allow probing optically thin samples, such as cellular membrane. Experimental approaches able to overcome this drawback are based on the use of metallic nanostructures, which lead to a huge amplification of the Raman yields thanks to the excitation of localized surface plasmon resonances. Surface-enhanced Raman scattering (SERS) and tip-enhanced Raman scattering (TERS) are examples of such innovative techniques, in which metallic nanostructures are assembled on a flat surface or on the tip of a scanning probe microscope, respectively. Herein, we provide a couple of examples (red blood cells and bacterial spores) aimed at studying cell membranes with these techniques.

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“…So far, RS has been widely employed in an impressive number of studies of in vitro and in vivo biological systems (DNA, peptide and proteins, microbes and virus, cells, etc.) [3][4][5]. Medical applications of RS are also very numerous and have been described in review papers and textbooks [6][7][8][9][10][11].…”

Section: Introductionmentioning

confidence: 99%

Raman-spectroscopy-based biosensing for applications in ophthalmology

Rusciano

Capriglione

Pesce

et al. 2013

Optical Sensors 2013

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Cell-based biosensors rely on the detection and identification of single cells as well as monitoring of changes induced by interaction with drugs and/or toxic agents. Raman spectroscopy is a powerful tool to reach this goal, being non-destructive analytical technique, allowing also measurements of samples in aqueous environment. In addition, micro-Raman measurements do not require preliminary sample preparation (as in fluorescence spectroscopy), show a finger-print spectral response, allow a spatial resolution below typical cell sizes, and are relatively fast (few s or even less). All these properties make micro-Raman technique particularly promising for high-throughput on-line analysis integrated in lab-on-a-chip devices. Herein, we demonstrate some applications of Raman analysis in ophthalmology. In particular, we demonstrate that Raman analysis can provide useful information for the therapeutic treatment of keratitis caused by Acanthamoeba Castellanii (A.), an opportunistic protozoan that is widely distributed in the environment and is known to produce blinding keratitis and fatal encephalitis. In particular, by combining Raman analysis with Principal Component Analysis (PCA), we have demonstrated that is possible to distinguish between live and dead cells, enabling, therefore to establish the effectiveness of therapeutic strategies to vanquish the protozoa. As final step, we have analyzed the presence of biochemical differences in the conjunctival epithelial tissues of patients affected by keratitis with respect to healthy people. As a matter of facts, it is possible to speculate some biochemical alterations of the epithelial tissues, rendering more favorable the binding of the protozoan. The epithelial cells were obtained by impression cytology from eyes of both healthy and keratitis-affected individuals. All the samples were analyzed by Raman spectroscopy within a few hours from cells removal from eyes. The results of this analysis are discussed.

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Label-Free Probing of G-Quadruplex Formation by Surface-Enhanced Raman Scattering

Cited by 54 publications

References 41 publications

Qualitative SERS analysis of G-quadruplex DNAs using selective stabilising ligands

Qualitative SERS analysis of G-quadruplex DNAs using selective stabilising ligands

Cell Imaging by Spontaneous and Amplified Raman Spectroscopies

Raman-spectroscopy-based biosensing for applications in ophthalmology

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