Near-infrared nucleic acids sensing and imaging using lanthanide-based nanoparticles capped with DNA

Project Information

Upbiosens

Grant agreement ID: 794410

DOI

10.3030/794410

Project closed

EC signature date 14 March 2018

Start date 1 May 2019

End date 30 April 2021

Funded under

• EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions

Total cost

€ 185 076,00

EU contribution

€ 185 076,00

Coordinated by

CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE CNRS
France

Periodic Reporting for period 1 - Upbiosens (Near-infrared nucleic acids sensing and imaging using lanthanide-based nanoparticles capped with DNA)

Reporting period: 2019-05-01 to 2021-04-30

Summary of the context and overall objectives of the project

Biosensing refers to the use of a biological derived or bioinspired receptor to detect the presence of a particular analyte with high specificity. The development of highly sensitive and specific nucleic acids (NA), i.e. DNA or RNA, biosensors has received increasing attention due to their crucial role in diagnostic and therapeutic applications (e.g. detection of disease biomarkers). Especially, the development of multiplexed micro-RNA sensors (miRNA, small non-coding RNAs) due to the role of miRNAs as regulators of gene expression. Overexpression levels can be associated with human diseases such as cancer and today, miRNAs are so-called next-generation biomarkers. Despite the development of methods to analyze NA, such as polymerase chain reaction (PCR), some technical shortcomings still hinder an easy-to-use, reproducible, storable, rapid, sensitive, specific, versatile, and multiplexed NA detection.
Due to the near-infrared (NIR) excitation, multiple and narrow emission bands over a broad wavelength range, and lack of photoblinking and photobleaching, the use of upconversion nanoparticles (UCNPs) offers the opportunity to design highly sensitive multiplexed UCNPs-based sensing probes.
The Upbiosens project describes a novel method to produce a nanobiosensor combining the sensing ability of the DNA strands and the optical properties of the UCNPs as the transducer element to detect and visualize DNA or/and miRNA. Hence, the project addresses the problem of capping UCNPs with DNA while keeping their hybridization capacity as well as controlling the UCNP size, overcoming one of the main limitations of the UCNPs for their application as NA sensors. The outcome of the project can be promisingly bio-implemented, such as in prognosis, diagnostics, and treatment of diseases, gene therapy, or forensic analysis.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

The Upbiosens project was carried out at Centre National de la Recherche Scientifique (CNRS), first in the Institut de Biologie Intégrative de la Cellule (I2BC, Orsay) and from September 2019 in the Laboratoire de Chimie Organique, Bioorganique, Réactivité et Analyse (COBRA, Mont-Saint-Aignan). The work performed during the financed period can be divided into three main axes: i) Developing a new synthesis method for obtaining stable DNA capped UCNPs, ii) Set-up a Förster Resonance Energy Transfer (FRET)-based homogeneous hybridization assay for detection of NA in solution, and iii) NA imaging in living cells. The results of the studies are published in two peer-reviews journals which are freely accessible through the open access repository Zenodo (https://zenodo.org/). Forthcoming publications will be also available in that repository. The results of the Upbiosens project have been also presented in four international conferences (SFNano-CNano joint meeting, 2019, Dijon; ANNIC 2019, Paris; IV Jornada de la mujer investigadora, 2020, València; UPCONline 2021).

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

The Upbiosens project has mainly addressed the synthesis of stable DNA-functionalized UCNPs while keeping their hybridization capacity as well as controlling the UCNP size. The DNA functionalization method developed within the project is versatile since it can be extended to different NA sequences, i.e. DNA or RNA, and, consequently, the nanosensors can be applied to many DNA and miRNA biomarkers.
This novel technology can be integrated into a broad range of applications, such as gene therapy, diagnosis/therapy of genetic diseases as well as prognosis, diagnosis, and treatment of diseases. Thus, the outcome of the project may have a potential impact on the understanding of the causes and mechanisms of genetic diseases as well as the ability to monitor, detect and treat diseases. Likewise, those results will allow the development of advanced technology to construct a currently unavailable simple-to-use but high-performance imaging tool based on UCNPs for NA sensing and imaging.
Additionally, the project has contributed to the understanding of the energy transfer mechanisms in UC PL processes in UCNPs, as well as to the mechanisms involved in FRET processes from UCNPs to an energy acceptor upon NIR excitation, which are not yet completely elucidated and is crucial for the wide and successful application of UC luminescent materials as sensors.

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