CRISPR-Clear: A Fieldable Detection Procedure for Potential CRISPR-Cas9 Gene Drive Based Bioweapons
semanticscholar(2019)
摘要
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.
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