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CRISPR-Clear: A Fieldable Detection Procedure for Potential CRISPR-Cas9 Gene Drive Based Bioweapons

preprint
submitted on 22.02.2019 and posted on 22.02.2019 by Anna C. Nieuwenweg, Martijn M. van Galen, Angelina Horsting, Jorrit W. Hegge, Aldrik Velders, Vittorio Saggiomo

Rapid progression in genetic modification research has made gene editing increasingly cheaper and easier to use. CRISPR-Cas9 for example, allows for the specific alteration of the genome of an organism with relative simplicity and low costs. This raised a worrying question; can genetic modification techniques be used to create novel bioweapons? A specific scenario is the initiation of a synthetic gene drive for malicious purposes. A synthetic gene drive can be used to quickly spread a mutation through an entire population. This mutation could alter vectors in such a way that they will spread human diseases or eradicate essential organisms. Since a gene drive spreads efficiently through a population, timely detection is essential. Thus, a quick and field deployable screening method is needed to counteract the malicious use of gene drives.

Here, we show a battery-operated, sensitive screening method, named CRISPR-Clear, for the detection of gene drive modified organisms. CRISPR-Clear is based on the combination of three components: 1) A DNA amplification technique known as loop-mediated isothermal amplification (LAMP) for detecting the presence of a gene drive; b) a portable battery-operated Arduino device which heats up the sample to allow DNA amplification, and c) a naked-eye visualization of the results.

We designed and tested six LAMP primers targeting a Cas9 endonuclease-based gene drive, assembled a battery-operated Arduino device and tested the naked-eye visualization method. In addition, we were able to detect the presence of the Cas9 gene, extracted from a transformed bacteria, providing a proof-of-concept of the CRISPR-Clear device.

Funding

NWO - Topsector Student competition 2017

History

Email Address of Submitting Author

vittorio.saggiomo@wur.nl

Institution

Wageningen University and Research

Country

Netherlands

ORCID For Submitting Author

0000-0001-7196-602X

Declaration of Conflict of Interest

No conflict of interests

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