<i>Drosophila melanogaster</i>as a Model for Gene Drive Systems

Champer, Jackson

doi:10.1079/9781800621176.0009

Cited by 2 publications

(3 citation statements)

References 85 publications

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“…Several gene-drive based versions of these strategies require the use of regulatory sequences that drive expression primarily or exclusively in the germline, particularly the ones that utilize engineered site-specific homing endonucleases. When expression of an endonuclease such as Cas9 is activated in the male and/or female germline in a hemizygote, cleavage of the target site on the wild-type chromosome followed by homology-directed DNA repair results in an increase in the frequency of the drive transgene in the population [47].…”

Section: Germline-specific Promotersmentioning

confidence: 99%

Use of Insect Promoters in Genetic Engineering to Control Mosquito-Borne Diseases

Bottino-Rojas

James

2022

Biomolecules

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Mosquito transgenesis and gene-drive technologies provide the basis for developing promising new tools for vector-borne disease prevention by either suppressing wild mosquito populations or reducing their capacity from transmitting pathogens. Many studies of the regulatory DNA and promoters of genes with robust sex-, tissue- and stage-specific expression profiles have supported the development of new tools and strategies that could bring mosquito-borne diseases under control. Although the list of regulatory elements available is significant, only a limited set of those can reliably drive spatial–temporal expression. Here, we review the advances in our ability to express beneficial and other genes in mosquitoes, and highlight the information needed for the development of new mosquito-control and anti-disease strategies.

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Section: Germline-specific Promotersmentioning

confidence: 99%

Use of Insect Promoters in Genetic Engineering to Control Mosquito-Borne Diseases

Bottino-Rojas

James

2022

Biomolecules

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“…HEG (I-SceI) - based homing gene drives have been successfully performed in Anopheles gambiae and Drosophila melanogaster ( 12 – 15 ). Since it proved difficult in practice to adapt HEGs to target essential genes, the advent of easily programmable “RNA-guided endonucleases” such as Cas9 has expedited gene drive research tremendously, showing the potential of this approach to genetically alter or suppress wild populations of insect pests ( 16 – 18 ).…”

mentioning

confidence: 99%

“…Once the target is recognized, Cas9 binds to its sequence and promotes DSBs. CRISPR/Cas9-based homing gene drive systems have been demonstrated in a wide range of organisms including yeast ( 21 ), D. melanogaster ( 18 , 22 ), mice ( 23 ), and the mosquito disease vectors A. gambiae ( 24 ), A. stephensi ( 25 ), and Aedes aegypti ( 26 ). The systems typically contain three elements, a Cas9 gene driven by a promoter active in the germline, one or more sgRNA genes controlled by a U6 gene promoter, and a fluorescent protein marker gene to monitor biased inheritance ( 24 , 25 ).…”

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confidence: 99%

CRISPR/Cas9-based split homing gene drive targeting doublesex for population suppression of the global fruit pest Drosophila suzukii

Yadav

Butler

Yamamoto

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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Genetic-based methods offer environmentally friendly species-specific approaches for control of insect pests. One method, CRISPR homing gene drive that target genes essential for development, could provide very efficient and cost-effective control. While significant progress has been made in developing homing gene drives for mosquito disease vectors, little progress has been made with agricultural insect pests. Here, we report the development and evaluation of split homing drives that target the doublesex ( dsx ) gene in Drosophila suzukii , an invasive pest of soft-skinned fruits. The drive component, consisting of dsx single guide RNA and DsRed genes, was introduced into the female-specific exon of dsx , which is essential for function in females but not males. However, in most strains, hemizygous females were sterile and produced the male dsx transcript. With a modified homing drive that included an optimal splice acceptor site, hemizygous females from each of the four independent lines were fertile. High transmission rates of the DsRed gene (94 to 99%) were observed with a line that expressed Cas9 with two nuclear localization sequences from the D. suzukii nanos promoter. Mutant alleles of dsx with small in-frame deletions near the Cas9 cut site were not functional and thus would not provide resistance to drive. Finally, mathematical modeling showed that the strains could be used for suppression of lab cage populations of D. suzukii with repeated releases at relatively low release ratios (1:4). Our results indicate that the split CRISPR homing gene drive strains could potentially provide an effective means for control of D. suzukii populations.

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Drosophila melanogasteras a Model for Gene Drive Systems

Cited by 2 publications

References 85 publications

Use of Insect Promoters in Genetic Engineering to Control Mosquito-Borne Diseases

Use of Insect Promoters in Genetic Engineering to Control Mosquito-Borne Diseases

CRISPR/Cas9-based split homing gene drive targeting doublesex for population suppression of the global fruit pest Drosophila suzukii

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