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Nucleic Acid detection with Short Prokaryotic Argonautes (NASPA)

Project description

Next-gen rapid diagnostics

The COVID-19 pandemic created a demand for swift, flexible diagnostic devices. Identifying particular genetic sequences is necessary for pathogen identification, mutation tracking, and the diagnosis of genetic disorders. Conventional PCR assays are accurate but need to be carried out in specialised laboratories by trained personnel, thus preventing their use in clinics or the field. In this context, the ERC-funded NASPA project aims to use short prokaryotic Argonautes and synthetic guide RNAs. NASPA can detect virtually any DNA or RNA sequence in a single reaction, producing a fluorescent glow upon detection. With point-of-care and field deployment in mind, NASPA endeavours to provide sensitive, rapid, and flexible diagnostics, bridging the gap between lab accuracy and real-world availability.

Objective

The SARS-CoV-2 pandemic highlighted the need for versatile diagnostics tools. Detecting presence of specific nucleic acid sequences is a powerful method for pathogen diagnostics, phenotyping, and determination of single nucleotide polymorphisms or other mutations underlying genetic diseases. PCR-based methods, the gold standard in nucleic acid detection, require specialized equipment and trained personnel, limiting its applicability at point-of-care or in-field locations. Alternative isothermal amplification/CRISPR-Cas-based detection methods suffer from technical limitations and a crowded patent landscape.

Nucleic Acid detection with Short Prokaryotic Argonautes (NASPA) is a versatile, sensitive, and specific one-pot nucleic acid detection method that can be used for point-of-care or in-field diagnostics. NASPA builds on our research in which we characterized short prokaryotic Argonaute (pAgo) systems. In NASPA, short pAgos are programmed with synthetic 15-21 nt guide RNAs to detect complementary DNA sequences. Upon detection of the DNA, breakdown of reporter ε-NAD+ is triggered, resulting in a traceable fluorescent signal. As there are no sequence requirements for the guide RNA, and guide RNAs can readily be synthesized, NASPA facilitates detection of virtually any sequence of choice.

We have proof of concept for NASPA-based diagnostics in a laboratory setting (technology readiness level (TRL) 3), and intellectual property has been protected through patent applications. The proposed ERC PoC project has two main objectives 1) enhance NASPA and enable detection of RNA and DNA sequences of economic and/or medical relevance in point-of-care and in-field settings, reaching TLR 5, and 2) build a network of stakeholders including academic and commercial partners to expedite the development of NASPA-based applications and establish a valorisation strategy. Achieving these objectives will pave the way for further development and valorisation of NASPA-based diagnostics.

Fields of science (EuroSciVoc)

Programme(s)

Topic(s)

Funding Scheme

HORIZON-ERC-POC - HORIZON ERC Proof of Concept Grants

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Call for proposal

(opens in new window) ERC-2024-POC

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Host institution

WAGENINGEN UNIVERSITY
Net EU contribution
€ 150 000,00
Address
DROEVENDAALSESTEEG 4
6708 PB Wageningen
Netherlands
Region
Oost-Nederland Gelderland Veluwe
Activity type
Higher or Secondary Education Establishments
Links
Total cost
No data

Beneficiaries (1)