2023
DOI: 10.1101/2023.01.27.525922
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Engineered Antiviral Sensor Targets Infected Mosquitoes

Abstract: Escalating vector disease burdens pose significant global health risks, so innovative tools for targeting mosquitoes are critical. We engineered an antiviral strategy termed REAPER (vRNA Expression Activates Poisonous Effector Ribonuclease) that leverages the programmable RNA-targeting capabilities of CRISPR Cas13 and its potent collateral activity. Akin to a stealthy Trojan Horse hiding in stealth awaiting the presence of its enemy, REAPER remains concealed within the mosquito until an infectious blood meal i… Show more

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Cited by 7 publications
(1 citation statement)
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“…To generate a rapid assay for transgene detection that could be used alongside future pgSIT field trials, we adapted the sensitive enzymatic nucleic acid sequence reporter (SENSR) (Brogan et al, 2021a) to rapidly detect pgSIT DNA fragments in Cas9 and gRNA dsx,ix, β Tub constructs in mosquitoes. In SENSR, the presence of transgenic sequences triggers the activation of the Ruminococcus flavefaciens CasRx/gRNA collateral activity that cleaves a quenched probe, resulting in a rapid fluorescent readout (Brogan et al, 2021b; Dalla Benetta et al, 2023) (Brogan et al, 2021a). Multiple ratios of trans-hemizygous pgSIT to WT mosquitoes - 0:1; 1:1; 1:5; 1:10; 1:25; 1:50 and a no template control (NTC) were used to challenge SENSR.…”
Section: Resultsmentioning
confidence: 99%
“…To generate a rapid assay for transgene detection that could be used alongside future pgSIT field trials, we adapted the sensitive enzymatic nucleic acid sequence reporter (SENSR) (Brogan et al, 2021a) to rapidly detect pgSIT DNA fragments in Cas9 and gRNA dsx,ix, β Tub constructs in mosquitoes. In SENSR, the presence of transgenic sequences triggers the activation of the Ruminococcus flavefaciens CasRx/gRNA collateral activity that cleaves a quenched probe, resulting in a rapid fluorescent readout (Brogan et al, 2021b; Dalla Benetta et al, 2023) (Brogan et al, 2021a). Multiple ratios of trans-hemizygous pgSIT to WT mosquitoes - 0:1; 1:1; 1:5; 1:10; 1:25; 1:50 and a no template control (NTC) were used to challenge SENSR.…”
Section: Resultsmentioning
confidence: 99%