CRISPR-based gene drives generate super-Mendelian inheritance in the disease vectorCulex quinquefasciatus

Abstract

Culexmosquitoes pose a significant public health threat as vectors for a variety of diseases including West Nile virus and lymphatic filariasis, and transmit pathogens threatening livestock, companion animals, and endangered birds. Rampant insecticide resistance makes controlling these mosquitoes challenging and necessitates the development of new control strategies. Gene drive technologies have made significant progress in other mosquito species, although similar advances have been lagging inCulex. Here we test the first CRISPR-based homing gene drive forCulex quinquefasciatus, demonstrating the possibility of using this technology to controlCulexmosquitoes. Our results show that the inheritance of two split-gene-drive transgenes, targeting different loci, are biased in the presence of a Cas9-expressing transgene although with modest efficiencies. Our findings extend the list of disease vectors where engineered homing gene drives have been demonstrated to includeCulexalongsideAnophelesandAedes, and pave the way for future development of these technologies to controlCulexmosquitoes.

Germline transformation of the West Nile Virus and avian malaria vectorCulex quinquefasciatusSay using the piggyBac transposon system

“…Transgenic Cx. quinquefasciatus have previously been generated using CRISPR-Cas9 mediated HDR (Feng et al, 2021; Harvey-Samuel et al, 2023; Purusothaman et al, 2021) and the Hermes transposable element (Allen et al, 2001; Allen and Christensen, 2004), however to date there are no reports of successful transformation using the PB system. This is surprising, given the broad species range, including yeast, plants, insects and mammals (Ding et al, 2005; Gregory et al, 2016; Nishizawa-Yokoi and Toki, 2021; Zhu et al, 2018), and previous success using PB in other mosquito genera (Grossman et al, 2001; Labbé et al, 2010a; Lobo et al, 2002; Nolan et al, 2002; Perera et al, 2002).…”

Gene drive and genetic sex conversion in the global agricultural pest Ceratitis capitata

“…Homing gene drives were originally proposed 20 years ago 1 are now under development in multiple insect species of medical, agricultural, or ecological importance [2][3][4][5][6][7][8][9][10][11][12][13] . Suppressive homing gene drives are designed to reduce the population size of harmful pest organisms by, in most cases, targeting genes essential for insect fitness using a homing CRISPR endonuclease expressed in the germline.…”

Gene drive and genetic sex conversion in the global agricultural pestCeratitis capitata

“…Homing gene drives were originally proposed 20 years ago 1 are now under development in multiple insect species of medical, agricultural, or ecological importance [2][3][4][5][6][7][8][9][10][11][12][13] . Suppressive homing gene drives are designed to reduce the population size of harmful pest organisms by, in most cases, targeting genes essential for insect fitness using a homing CRISPR endonuclease expressed in the germline.…”